猪传染性胃肠炎M基因的克隆及序列分析
摘要
猪传染性胃肠炎病毒(Transmissible Gastroenteritis Virus, TGEV)属于冠状病毒
科冠状病毒属，其可导致猪传染性胃肠炎。对养猪业危害性极大。抗毒素和抗生素等常
规药物对本病无任何治疗作用，预防该病的有效方法是疫苗接种。本研究以猪传染性胃
肠炎病毒感染的乳猪小肠病料为材料，自行设计两对引物，通过套式（RT-nested）PCR
及普通的RT-PCR扩增出猪传染性胃肠炎病毒M基因全序列，经测序、同源性比较后，进
行深入的分析，主要研究结果如下：
1．本试验以乳猪小肠组织病料为材料，用Trizol法提取总RNA，经甲醛凝胶变性
电泳，能清晰的看到28s、18srRNA两条亮带以及一条较弱的5srRNA带。结果表明，所提
取的RNA完整性好，没有发生降解，完全可以用于RT-PCR反应。
2．根据GeneBank中发表的猪传染性胃肠炎病毒M基因序列，利用软件设计两对引物
（外引物和内引物），在普通的RT- PCR方法基础上，建立了 RT-nested PCR体系，扩
增出猪传染性胃肠炎病毒M基因的全序列，使用内外引物扩增的片段长度分别为1037bp、
1328bp 。结果表明，用双引物的RT-nested PCR较普通的RT- PCR扩增方法更具特异性，
重复性好，能很好的避免病毒检测时假阳性的出现。且较传统的方法如电镜技术、免疫
荧光技术、ELISA等更方便可靠，完全可以作为一种新型的、特异性强的病毒检测的方
法。
3．利用 RT- PCR 方法，扩增出猪传染性胃肠炎病毒 M 基因全序列，长度为 1 328 bp，
扩增产物经回收、转化,克隆在 pGEM-T Easy 载体的 T 位点上。用 EcoRⅠ酶切鉴定重组
质粒 DNA，挑选出阳性克隆菌，进行序列测定后。利用 CLUSTAL（1.82）软件进行同源
性分析。结果显示，扩增的猪传染性胃肠炎病毒 M 基因序列与 GeneBank 中登录号为
AY587883 和 AF302262 的猪传染性胃肠炎病毒 M 基因的核苷酸序列同源性分别为 97.2%，
97.3%。氨基酸同源性分别为 97.3%，98.0％。软件分析表明，各有 22 处和 21 处碱基发
生了突变，其中一处碱基发生了缺失，7 个和 5 个氨基酸发生了转换。
本研究不仅为对TGEV进行深入的分子生物学研究提供了资料，为其基因工程疫苗
的研制奠定了基础，并且也为该病毒的鉴定提供了一个便捷有效的方法，对猪传染性胃
肠炎的防治有重要的现实意义。
Porcine transmissible gastroenteritis virus (TGEV) belongs to the family coronaviridae.
The virus causes transmissible gastroenteritis (TGE) which does very much harm to the pig
farms. The normal medicines such as antibiotic and antitoxins have no effect to the disease.
Vaccination is the effective means for the prevention of TGE.The study isolated RNA from
enteritis tissue. Two pairs of primes were designed to amplify cDNA sequence of M gene by a
Transcription-nested Polymerase chain Reaction (RT-PCR)and a Reverse Transcription-nested
Polymerase chain Reaction (RT-nested PCR) . The products of PCR were sequenced and
homology compared .Then they were analysed deeply . The following was the main
experiments and results:
1. The study isolated RNA from enteritis tissue by Trizol method. After formaldehyde
gel denaturation electrophoresis, we can see clearly two light bands(28s and 18srRNA) and a
vague band(5srRNA).The result indicated that the integrality of the isolation RNA was very
good and hardly was degradated.It may be used in RT-PCR.
2. According to the published cDNA sequence of TGEV M gene, with Prime5.0 two
pairs of primes were designed to amplify the products which were 1037 bp and 1328 bp
respectively by RT-nest PCR based on common RT-PCR. The result demonstrated that the
method has higher specialist and repeation. It can detect correctly virus and avoid false
positivism. Compared with tranditional methods, such as electron microscopy and
immunization fluorescence technology and ELISE, it is more convenience and
credibility .The results showed that RT-nest PCR may be one method which is neoteric and
reliable to detect the RNA virus especially.
3. The assay amplified the full seqence of M gene which is 1 328 bp nucleotide acids by
RT-PCR . The amplified product was extracted ， transformed and cloned into pGEM-T
Easy vector. The recombinant plasmid was tested by restricted digestion of EcoRⅠand the
sequencing. The sequence analysis indicated that the sequence homology of nucleotide acids
was 97.2%, 97.3% and the sequence homology of amino acid was 97.3%,98.0% which were
analyzed by CLUSTUL(1.82) when it was compared with corresponding sequencing of the
different strains in GeneBank. There were twenty-four nucleotide acids mutation and seven
amino acids mutation. And the further analysis indicated that these mutations of nucleotide
acids were no meaning and did not affect the fuction of M protein.
The study provided not only the important basis for further research of TGEV molecular
properties and the preparation of vaccine, but also an useful viral strain discrimination method.
In brief, it provided vital practical value for molecular study of TGEV , development of
vaccine and prevention of TGE as well.
科冠状病毒属，其可导致猪传染性胃肠炎。对养猪业危害性极大。抗毒素和抗生素等常
规药物对本病无任何治疗作用，预防该病的有效方法是疫苗接种。本研究以猪传染性胃
肠炎病毒感染的乳猪小肠病料为材料，自行设计两对引物，通过套式（RT-nested）PCR
及普通的RT-PCR扩增出猪传染性胃肠炎病毒M基因全序列，经测序、同源性比较后，进
行深入的分析，主要研究结果如下：
1．本试验以乳猪小肠组织病料为材料，用Trizol法提取总RNA，经甲醛凝胶变性
电泳，能清晰的看到28s、18srRNA两条亮带以及一条较弱的5srRNA带。结果表明，所提
取的RNA完整性好，没有发生降解，完全可以用于RT-PCR反应。
2．根据GeneBank中发表的猪传染性胃肠炎病毒M基因序列，利用软件设计两对引物
（外引物和内引物），在普通的RT- PCR方法基础上，建立了 RT-nested PCR体系，扩
增出猪传染性胃肠炎病毒M基因的全序列，使用内外引物扩增的片段长度分别为1037bp、
1328bp 。结果表明，用双引物的RT-nested PCR较普通的RT- PCR扩增方法更具特异性，
重复性好，能很好的避免病毒检测时假阳性的出现。且较传统的方法如电镜技术、免疫
荧光技术、ELISA等更方便可靠，完全可以作为一种新型的、特异性强的病毒检测的方
法。
3．利用 RT- PCR 方法，扩增出猪传染性胃肠炎病毒 M 基因全序列，长度为 1 328 bp，
扩增产物经回收、转化,克隆在 pGEM-T Easy 载体的 T 位点上。用 EcoRⅠ酶切鉴定重组
质粒 DNA，挑选出阳性克隆菌，进行序列测定后。利用 CLUSTAL（1.82）软件进行同源
性分析。结果显示，扩增的猪传染性胃肠炎病毒 M 基因序列与 GeneBank 中登录号为
AY587883 和 AF302262 的猪传染性胃肠炎病毒 M 基因的核苷酸序列同源性分别为 97.2%，
97.3%。氨基酸同源性分别为 97.3%，98.0％。软件分析表明，各有 22 处和 21 处碱基发
生了突变，其中一处碱基发生了缺失，7 个和 5 个氨基酸发生了转换。
本研究不仅为对TGEV进行深入的分子生物学研究提供了资料，为其基因工程疫苗
的研制奠定了基础，并且也为该病毒的鉴定提供了一个便捷有效的方法，对猪传染性胃
肠炎的防治有重要的现实意义。
Porcine transmissible gastroenteritis virus (TGEV) belongs to the family coronaviridae.
The virus causes transmissible gastroenteritis (TGE) which does very much harm to the pig
farms. The normal medicines such as antibiotic and antitoxins have no effect to the disease.
Vaccination is the effective means for the prevention of TGE.The study isolated RNA from
enteritis tissue. Two pairs of primes were designed to amplify cDNA sequence of M gene by a
Transcription-nested Polymerase chain Reaction (RT-PCR)and a Reverse Transcription-nested
Polymerase chain Reaction (RT-nested PCR) . The products of PCR were sequenced and
homology compared .Then they were analysed deeply . The following was the main
experiments and results:
1. The study isolated RNA from enteritis tissue by Trizol method. After formaldehyde
gel denaturation electrophoresis, we can see clearly two light bands(28s and 18srRNA) and a
vague band(5srRNA).The result indicated that the integrality of the isolation RNA was very
good and hardly was degradated.It may be used in RT-PCR.
2. According to the published cDNA sequence of TGEV M gene, with Prime5.0 two
pairs of primes were designed to amplify the products which were 1037 bp and 1328 bp
respectively by RT-nest PCR based on common RT-PCR. The result demonstrated that the
method has higher specialist and repeation. It can detect correctly virus and avoid false
positivism. Compared with tranditional methods, such as electron microscopy and
immunization fluorescence technology and ELISE, it is more convenience and
credibility .The results showed that RT-nest PCR may be one method which is neoteric and
reliable to detect the RNA virus especially.
3. The assay amplified the full seqence of M gene which is 1 328 bp nucleotide acids by
RT-PCR . The amplified product was extracted ， transformed and cloned into pGEM-T
Easy vector. The recombinant plasmid was tested by restricted digestion of EcoRⅠand the
sequencing. The sequence analysis indicated that the sequence homology of nucleotide acids
was 97.2%, 97.3% and the sequence homology of amino acid was 97.3%,98.0% which were
analyzed by CLUSTUL(1.82) when it was compared with corresponding sequencing of the
different strains in GeneBank. There were twenty-four nucleotide acids mutation and seven
amino acids mutation. And the further analysis indicated that these mutations of nucleotide
acids were no meaning and did not affect the fuction of M protein.
The study provided not only the important basis for further research of TGEV molecular
properties and the preparation of vaccine, but also an useful viral strain discrimination method.
In brief, it provided vital practical value for molecular study of TGEV , development of
vaccine and prevention of TGE as well.
引文
[1] 何孔旺,林继煌,还红华,等 猪传染性胃肠炎病毒弱毒株的细胞培养特性及致病性
研究[J].中国兽医科技,2001,31(8):8-9.
[2] 王继科，刘常明，马思奇,等.猪传染性胃肠炎和猪流行性腹泻病毒免疫电镜的诊断
的研究[J].中国畜禽传染病,1991,2:22-25.
[3] 方定一. 兽医微生物学[M].北京：农业出版社,1979.
[4] 王明俊.兽医生物制品[M].北京：中国农业出版社,1996.
[5] 任晓峰.猪传染性胃肠炎病毒TH-98株S基因的克隆与鉴定.东北农业大学硕士学位
论文[J].2001
[6] 曹军平,朱爱萍,肖作焕,等.猪传染性胃肠炎的诊断与防制[J].中国动物检疫,
2000,6:29.
[7] 唐立杰，李一经.RT-PCR 方法快速诊断猪传染性胃肠炎[J].黑龙江畜牧兽
医,2001,12:26-27.
[8] 蔡宝祥.家畜传染病学[M].北京,中国农业出版社,2001,213-216.
[9] 周仲芳，立力复，罗长保,等.利用重组抗原建立间接酶联免疫吸附试验检测猪传染
性胃肠炎血清抗体[J].中国兽医杂志,2000,26 (10):12-14.
[10] 赵德明，张中秋，沈建中，主译.B. E.斯特劳，S. D.阿莱尔，W. L.蒙加林等,猪
病学[M],中国农业大学出版社,2000,9:263-276.
[11] 殷震，刘景华.动物病毒学[M].北京：科学出版社,1997,671-703.
[12] 冉智光,王玉春.猪传染性胃肠炎病毒分子生物学研究新进展[J].天津畜牧兽
医,1998,4:4-6.
[13] 佟有恩，冯力，李伟杰，等. 猪传染性胃肠炎与猪流行性腹泻二联弱毒疫苗的研
究[J]. 中国预防兽医学报，1999,21(6):406-410
[14] 师东方.猪轮状病毒和传染性胃肠炎病毒核酸免疫的研究[J]. 东北农业大学博士
学位论文. 2002.
[15] 孙树汉. 核酸疫苗[M]. 上海：第二军医大学出版社，1999.
[16] 梁雨等. 核酸免疫可特异性地诱发小鼠对乙型肝炎病毒表面抗原 S 的免疫反应
[J]。病毒学报，1996,12(3):193.
[17] 王军阳. 核酸疫苗的安全性及伦理学问题[J]。中国医学伦理学，2000,2: 54.
[18] 萨姆布鲁克 J, 弗里奇 E F, 曼尼阿蒂斯 T. 分子克隆实验指南[M]. 金冬雁，
黎孟枫，等. 第二版. 北京:科学出版社,1993,26,674-683.
[19] 邹勇, 钱永清. 猪流行性腹泻、猪传染性胃肠炎和猪轮状病毒 RT-PCR 鉴别诊断技
术研究[J]. 上海农业学报,2003,19(2):82-84.
[20] 吴凌, 李一经. 应用 RT-nested PCR 检测猪流行性腹泻病毒的研究[J]. 中国兽医
40 猪传染性胃肠炎病毒 M 基因的克隆及序列分析
科技,2003,33(3):27-30.
[21]乔宪凤，熊忠良，郑新民等.PEDV 膜蛋白基因 PT-PCR 最适反应条件的确立[J].湖北
畜牧兽医，2001，1，8-10.
[22] Saif L J,Wesley R D.Transmissible gastroenteritis In Diseases of Swine[M].
Iowa State University Press,Ames,IA,1992,362-386.
[23] Enjuanes L,Van der Zeijst B A M. Molecular Basis of Transmissible
Gastroenteritis Coronavirus Epidemiology[M].PlenumPress,New York,N.Y.
1995, 337-376.
[24] Paton D, Ibata G, Sands J, McGoldrick A.Detection of Transmissible
Gastroenteritis Virus by RT-PCR and Differentiation from Porcine
Respiratory Coronavirus[J].J Virol Methods,1997,66(2):303-309.
[25] Enjuanes L, Spaan W, Snijder E Cavanagh D. Nidovirales. Classification and
Nomenclature of Viruses In Virus taxonomy[M].New York,Academic Press,2000,
827-834.
[26] Wesley RD,Woods R and Cheuny A. Genetic Basis of the Pathogenesis of
Transmissible Gastroenteritis Virus[J].Journal of Virol,1990,64(10):
4761-4766.
[27] Paton D, Lowings P. Discrimination between Transmissible Gastroenteritis
virus Isolates[J].Arch Virol.1997b,142(8):1703-1711.
[28] Goodwin R F W ,Jennings A R.A Highly Infectious Gastroenteritis of Pig[J].
Vet Rec,1958,70:271-272.
[29] Sasahara J, Harada K, Hayashi S and Watanabe M.Studies on Transmissible
Gastroenteritis in Pig in Japan. Japan[J]. J. Vet.Sci,1958,20:1-6.
[30] Siddell S G. The Coronavifdae In The Viruses[M], New York,Plenum Press.
1995,418.
[31] Lai M C ,Cavanagh D. The Molecular Biology of Coronaviruses[J]. Advances
in Virus Research,1997,48:1-100.
[32] Caul E ,Egglestone S.Coronavirus in Humans In Virus Infections of the
Gastrointestinal Tract[M]. New York: Marcel Dekker,1982,179-193.
[33] Denison M R. In Viral Infections of the Respiratory Tract Edited by
Dolin[M]. New York,Marcel Dekker,1999,253-280.
[34] Enjuanes L, Sola I, Almazan F, Ortego J, Izeta A, Gonzalez J M, Alonso S,.
Coronavirus Derived Expression Systems[J].Journal of Biotechnology,2001，
88:183-204.
[35] Kemeny L J.Antibody Response in Pigs Inoculated with Transmissible
参考文献 41
Gastroenteritis Virus and Cross Reactions among Ten Isolates[J].Can J Comp
Med.1976,40: 209-214.
[36] Pederden M C, Ward J and Mengeling W L.Antigenic Relationship of Feline
Infectious Peritonitis Virus to Coronaviruses of Other Species[J].Arch.
Virol,1978,58:45-53.
[37] Reynolds D J, Garwes D J and Lucey S.Differentiation of Canine Coronavirus
and Porcine Transmissible Gastroenteritis Virus by Neutralization with
Canine，Porcine and Feline Sera[J].Vet. Microbiol,1980,5:283-290.
[38] Woods R D.Studies of Enteric Coronaviruses in a Feline Cell Line[J].Vet.
Microbiol,1982,7:427-435.
[39] Haeltennan E O.Epidemiological Studies of Transmissible Gastroenteritis
of Swine[J].Proc US Livest Sunit Assoc,1962,66:305-315.
[40] Klemm R C,Ristic M.The Effect of Propagation of Transmissible
Gastroenteritis(TGE) Virus in Pups and the Lung of Baby Pigs on the
Immunologic Properties of the VirusInt Congr Pig Vet Soc[M].Iowa State
Univ,1976,11.
[41] Pensaert M, Callebaut P and Vergote J.Isolation of a Porcine Respiratory,
Non-enteric Coronavirus Related to Transmissible Gastroenteritis[J].Vet
Q. 1986,8:257-261.
[42] Kim B, Kim O, Tai J H, Chae C.Transmissible Gastroenteritis Virus Induces
Apoptosis in Swine Testicular Cell Lines But not in Intestinal
Enterocytes[J].J Com Pathol,2000,123(1):64-66.
[43] Hooper B E and Haelterman E O.Concepts of Pathogenesis and Passive Immunity
in Transmissible Gastroenteritis of Swine[J].J Am Vet Med Assoc,
1966,149:1580-1586.
[44] Butler D G, Gall D G, Kelly M H,et al.Transmissible Gastroenteritis
Mechanisms Responsible for Diarrhea in an Acute Enteritis in Piglets[J].
J Clin Invest,1974,53:1335-1342.
[45] Comelius L M, Hooper B E and Haelterman E O. Changes in Fluid and Electrolyte
Balance in Baby Pigs with Transmissible Gastroenteritis[J].Am J Clin Pathol,
1968,2:105-113.
[46] Wagner J E, Beamer P D and Ristic M.Electron Microscopy of Intestinal
Epithelial Cells of Piglets Infected with a Transmissible Gastroenteritis
Virus[J].Can J Comp Med,1973,37:177-188.
[47] Dul G C, Ruckerbauer P,Transmissible Gastroenteritis Demonstration of the
42 猪传染性胃肠炎病毒 M 基因的克隆及序列分析
Virus from Field Specimens by Means of Cell Culture and Pig Inoculation[J].
Can J Comp Med,1977,41:357-363.
[48] Bohl E H. Diagnosis of Diarrhea in Pigs due to Transmissible Gastroenteritis
Virus or Rotavirus In Viral Enteritis in Humans and Animals[J].Ed Bricout
and Scherrer INSERM,1979,90:341-343.
[49] Stark S L, Fernelius A L, Booth G D et al.Transmissible gastroenteritis
(TGE) of swine Effect of Age of Swine Testes Cell Culture Monolayers on Plaque
Assays of TGE Virus[J].Can J Comp Med,1975,39:466-468.
[50] Black J W.Diagnosis of TGE by FA Evaluation of Accuracy on Field
Specimens[J].Proc US Anim Health Assoc,1971,75:492-498.
[51] Garwes D J, Stewart F, Cartwright S F et al. Differentiation of Porcine
Coronavirus from Transmissible Gastroenteritis Virus. Vet Rec. 1988,122:
86-87.
[52] Van Nieuwstadt A P and Pol J M A. Isolation of a TGE Virus-related Respiratory
Coronavirus Causing Fatal Pneumonia in Pigs[J].Vet Rec,1989,124:43-44.
[53] Benfield D A, Jackwood D J, Bae I,et al.Detection of Transmissible
Gastroenteritis Virus Using cDNA Probes[J].Arch Virol,1991,116:91-106.
[54] Paton D,Lowings P.Discrimination between Transmissible Gastroenteritis
virus Isolates[J].Arch Virol,1997,142(8):1703-1711.
[55] Van Cott J L, Brim T A, Simkins R A,et al.Antibody-secreting Cells to
Transmissible Gastroenteritis Virus and Porcine Respiratory Coronavirus
in Gut and Bronchus-associated Lymphoid Tissues of Neonatal Pigs[J].J
Immunol, 1993,150:3990-4000.
[56] Saif L J, Van Cott J L ,Brim T A.Immunity to Transmissible Gastroenteritis
Virus and Porcine Respiratory Coronavirus Infections in Swine[J].Vet
Immunol Immunopathol,1994,43:89-97.
[57] Anton I M, Gonzalez S, Bullido M J,et al.Cooperation Between Transmissible
Gastroenteritis Coronavirus (TGEV) Structural Proteins in the in vitro
Induction of Virus-specific Antibodies[J].Virus Res,1996,46:111-124.
[58] Siddle S G, Anderson R, Cavanagh D, Fujiwara K, Coronaviridae[J].
Intervirology,1983,20:181-189.
[59] Spaan W,Cavanagh D,Horizinek M C.Coronaviruses Structure and Gene
Expression[J].Journal of Gen Virol,1988,69:2939-2952.
[60] Britton P and Page D W. Sequence of the S Gene from a Virulent British Field
Isolate of Transmissible Gastroenteritis Virus[J].Journal of Virus Res,
参考文献 43
1990,18:71-80.
[61] Curtis K M, Yount B,Baric R S.Heterologous Gene Expression from
Transmissible Gastroenteritis Virus Replicon Particles[J]. Journal of
Virol,2002,76(3):1422-1434.
[62] Cavanagh D, Brian D A, Brinton M A et al. Revision of the Taxonomy of the
Coronaviruses Torovirus and Arterivirus Genera[J].Archive of Virol,1994,
135(2):227-237.
[63] Kevin W, Paul P, Britton ,Michael E G. Sequence Analysis of the Leader RNA
of Transmissible Gastroenteritis Virus[J].Virus Gene,1990,4:289-303.
[64] Hiscox J A.Characterization of the Transmissible Gastroenteritis Vinis
(TGEV) Transcription Initiation Sequence[J].Adv Exp Med Biol,1995,
380:529-535.
[65] Wesley R, Woods R and Cheuny, A. Nucleotide Sequence of the E2-peplomer
Protein Gene and Partial Nucleotide Sequence of Upstream Polymerase Gene
of Transmissible Gastroenteritis Virus[J].Adv Exp Med Biol,
1990,276:301-306.
[66] Paul B, Kevin W. The Cloning and Sequencing of the Virion Protein Genes
from a British Isolate of PRCV Comparison with TGEV[J]. Journal of Virus
Research, 1991,21:181-198.
[67] Julian A, Hiscox, David C et al.Quantification of Individual Subgenomic
mRNA Species during Replication of the Transmissible Gastroenteritis
Virus[J]. Virus Res,1995,36:119-130.
[68] Gebauer F, Posthumus W P, Correa 1, Sune C, Smerdou C.Residues Involved
in the Antigenic Sites of Transmissible Gastroenteritis Coronavirus S
Glycoprotein[J].Virology,1991,83(1):225-238.
[69] Chen C M, Cavanagh D,Britton P.Cloning and Sequencing of a 8.4kb Region
from the 3'End of a Taiwanese Virulent Isolate of the Coronavirus
Transmissible Gastroenteritis Virus[J].Virus Res,1995,38(1):83-89
[70] Ballesteros L, Sanchez C ,Enjuanes L.Two Amino Acid Changes at the
N-terminus of TGEV Spike Protein Result in the Loss of Enteric
Tropism[J].Journal of Virol,1997,227(2):378-388.
[71] Christine K, Graham D, Yolken R et al.Point Mutations in the S Protein
Connect the Sialic Acid Binding Activity with the Enteropathogenicity of
TGEV[J]. Journal of Virol,1997,71(4):3285-3287.
[72] Krempl C, Schultze B, Laude H,Healer G.Point Mutations in the S Protein
44 猪传染性胃肠炎病毒 M 基因的克隆及序列分析
Connect the Sialic Acid Binding Activity with Enteropathogenicity of
Transmissibl Gastroenteritis
Virus[J]JournalofVirol,1997,71(4):3285-3287
[73] Delmas B, Rasschaert D, Gtodet M, Gelfi J.Four Major Antigenic Site of the
Coronavirus Transmissible Gastroenteritis Virus are Located on the
Amino-terminal Half of Spike Glycoprotein S[J].Journal of Gen Virol,
1990,71:1313-1323.
[74] Laude H, Rasschaert D, Delmas B,et al.Molecular Biology of Transmissible
Gastroenteritis Virus[J].Vet Virol,1990,23:147-154.
[75] Gebauer F, Posthumus W P, Correa 1, Sune C, Smerdou C.Residues Involved
in the Antigenic Sites of Transmissible Gastroenteritis Coronavirus S
Glycoprotein[J].Virology,199183(1):225-238.
[76] Correa I, Jimenez G, Sune C, Bullido M J.Antigenic Structure of the E2
Glycoprotein from Transmissible Gastroenteritis Coronavirus[J].Virus Res,
1988,10:77-94.
[77] Tuboly T, Nagy E.Immunogenicity of the S Protein of Transmissible
Gastroenteritis Virus Expressed in Baculo virus[J].Arch Virol,
1994,137(2):55-67.
[78] Bernard S and Laude H.Site-specific Alteration of Transmissible
Gastroenteritis Virus Spike Protein Results in Markedly Reduced
Pathogenicity. Journal of Gen[J].Virol,1995,76:2235-2241.
[79] Christine K, Graham D, Yolken R,et al.Point Mutations in the S Protein
Connect the Sialic Acid Binding Activity with the Enteropathogenicity of
TGEV[J].Journal of Virol,1997,71(4):3285-3287.
[80] Laviada M D, Videgain S P, Moreno L M.Expression of Swine Transmissible
Gastroenteritis Virus Envelope Antigens on the Surface of Infected Cell:
Epitopes externally exposed[J].Virus Res,1990,16:247-254.
[81] Sune C, Jimenez G, Correa I.Mechanisms of Transmissible Gastroenteritis
Virus Coronavirus Neutralization[J].Virology,1990,177:559-569.
[82] Delmas B, Rasschaert D, Gtodet M.Four Major Antigenic Site of the
Coronavirus Transmissible Gastroenteritis Virus are Located on the
Amino-terminal Half of Spike Glycoprotein S[J].Journal of Gen
Virol,1990,71:1313-1323.
[83] Delmas B, Gelfi J.Antigenic Structure of Transmissible Gastroenteritis
Virus Domains in the Peplomer Glycoprotein[H].Journal of Virol,1986,67:
参考文献 45
1405-1418.
[84] Have P.Infection with a New Porcine Respiratory Coronavirus in Denmark:
Serologic Differentiation from Transmissible Gastroenteritis Virus Using
Monoclonal Antibodies[M].In Coronaviruses and Their Diseases Plenum Press,
New York,1990,435-439.
[85] Sanchez C, Jimenez G, Laviada M D.Antigenic Homology among Coronavirus
Related to Transmissible Gastroenteriti Virus[J].Virology,1990,174:414
[86] Kim L, Hayes J. Molecular Characterization and Pathogenesis of
Transmissible Gastroenteritis Coronavirus (TGEV) and Respiratory
CoronavirusRC Field IsolatesCo-circulatingina Swine Herd[J].Arch
Virology,2000,145:1133-1147.
[87] Simpkins R A, Weilnau P A.Antigenic Variation among Transmissible
Gastroenteritis Virus (TGEV) and Porcine Respiratory Coronavirus Strains
Detected with Monoclonal Antibodies to the S Protein of TGEV[J].Am. J.
Vet. Res,1992,53:1253-1258.
[88] Niemann H, Boschek B, Evans D.Post-translational Glycosylation of
Coronavirus Glycoprotein E1 Inhibition by Monensin[J].Eur Mol Biol
Organ,1982,1:1499-1504.
[89] Jimenez G, Correa I, Melgosa M P.Critical Epitopes in Transmissible
Gastroenteritis Virus Neutralization[J].Journal of Virol,1986,60:131
一 139.
[90] Jones T ，Shenk T.Transmissible Gastroenteritis Virus of pigs[J].Vet Rec,
1997,41(16):427-428.
[91] Jacobs L, Van Der Zeust B A M,Horzined M C. Characterization and Translation
of Transmissible Gastroenteritis Virus RNAs[J].Journal of Virol,1986,57:
1010-1015.
[92] Carlos M. Sanchez, Izeta A, Jose M. Targeted Recombination Demonstrates
that the Spike Gene of Transmissible Gastroenteritis Coronavirus is a
Determinant of Its Enteric Tropism and Virulence[J].Journal of
Virol,1999,73:7607-7618.
[93] Taguchi F ，Fleming J O. Comparison of Six Different Murine Coronavius JHM
Variants by Monoclonal Antibodies against the E2 Glycoprotein[J].
Virology,1989,169:223-235
[94] Collins A R, Knobler R L, Powell H .Monoclonal Antibodies to Murine Hepatitis
Virus-4(strain JHM)Define the Viral Glycoprotein Responsible for
46 猪传染性胃肠炎病毒 M 基因的克隆及序列分析
Attachment and Cell-cell Fusion[J].Virology,1982,119:358-371.
[95] Sturman L S, Richard C S,Murine K V. Proteolytic Cleavage of the E2
Glycoprotein of Murine Coronavirus by Trysin and Separation of Two
Different 90K Cleavage Fragments[J].Journal of Virol,1985,56:904-911.
[96] Ballesteros L, Sanchez C ,Enjuanes L.Two Amino Acid Changes at the
N-terminus of TGEV Spike Protein Result in the Loss of Enteric
Tropism[J].Journal of Virol,1997,227(2):378-388.
[97] Christine K, Graham D, Yolken R et al. Point Mutations in the S Protein
Connect the Sialic Acid Binding Activity with the Enteropathogenicity of
TGEV Journal of Virol. 1997,71(4):3285-3287.
[98] Motokawa K. Comparison of the Amino Acid Sequence and Phylogenetic Analysis
of the Peplomer Integral Membrane and Nucleocapsid Protein of Feline Canine
Porcine Corona virus[J].Journal of Microbiology and immunology.
1996,40(6):425-433.
[99]Rasschaert D ,Laude H. The Predicted Primary Structure of the Peplomer
Protein E2 of the Porcine Coronavirus Transmissible Gastroenteritis
Virus[J].Journal of Gen Virol,1987,68:1883-1890.
[100]Charley B ， Laude H. Induction of Alpha Interferon by Transmissible
Gastroenteritis is Coronavirus:Role of Transmembrane Glycoprotein
E1[J].Journal of Virol,1988,62:8-11.
[101]Laude H, Rasschaert D, Delmas B,et al.Molecular Biology of Transmissible
Gastroenteritis Virus[J].Vet Virol,1990,23:147-154.
[102]Garwes D, Pocock D H,Wllaszka T M.The Polypeptide Structure of
Transmissible Gastroenteritis Virus[J].Journal of Gen Vrol,1975,29:25-34.
[103]Motokawa K.Comparison of the Amino Acid Sequence and Phylogenetic Analysis
of the Peplomer Integral Membrane and Nucleocapsid Protein of Feline Canine
and Porcine Coronavirus[J].Journal of Microbiology and Immunology,
1996,40(6):425-433.
[104]Nakange K,Yamanouchi K,Fujiwara K.Protective Effect of Monoclonal
Antibodies on Lethal Mouse Hepatitis Infection in Mice[J].Journal of Virol,
1986,59:168-171.
[105]Compton J，Rogers D B, Holmes K V, Fersch D.In vitro Replication of Mouse
Hepatitis Virus Strain A59[J].Journal of Virol,1987,61:1814-1820.
[106]Baric R S, Nelson G W J O, Deans R J.Interaction Between Coronavirus
Nucleocapsid Protein and Viral RNA Implications for Viral Transcription[J].
参考文献 47
Journal of Virol,1988,62:4280-4287.
[107]Stohlman L M, Baric R, Nelson G N, Welter L M .Specific Interaction between
Coronavirus Leader RNA and Nucleocapsid Protein[J].Journal of Virol,
1988,62:4288-4295.
[108]Boursnell M E G, Brown T D K, Foulds I J, Green P F. Completion of the Sequence
of the Genome of the Coronavirus Avian Infectious Bronchitis Virus[J].
Journal of Gen Virol,1987,68: 53-77.
[109]Wesley R D,Woods R,Cheuny A.Genetic Basis of the Pathogenesis of
Transmissible Gastroenteritis Virus[J]. Journal of Virol,1990,64(10):
4761-4766.
[110]Wesley R D, Woods R D ,Cheung A K.Genetic Analysis of Porcine Respiratory
Coronavirus an Attenuated Variant of Transmissible Gastroenteritis
Coronaviurs[J].Journal of Virol,1991,65:3369-3373.
[111]Julian A, Hiscox, David C ,et al.Quantification of Individual Subgenomic
mRNA Species during Replication of the Transmissible Gastroenteritis
Virus[J].Virus Res,1995,36:119-130.
[112]Leman A D, Straw B, Glock R D,et al.Disease of Swine[M] .Ames Iowa state
university press,1992:362-386.
[113] Garwes D J,Lucas M H,Higging D A,Pike B V ，Cratwright S F. Antigenicity
of Structural Components from Porcine Transmissible Gastroenteritis
Virus[J]. Vet Microbiol，1978,1979,3:179-190.
[114]Gough P M,Frank C J,Moore D G，et al. Lactogenic Immunity to Transmissible
Gastroenteritis Virus Induced by a Subunit Immunogen[J].Vaccine ，
1983,1:37-41.
[115]Posthumus W P,Enstra J A, Schaaper W M ，et al. Analysis and Simulation
of Neutralizing Epitope of Transmissible Gastroenteritis Virus[J]. J
Virol，1990,64:3304-3309.
[116]Castilla J,Sola I,Pintado B,Sanchez-Morgado J M，Enjuanes L. Lactogenic
Immunity in Transgenic Mice Producing Recombinant Antibodies Neutralizing
Coronavirus[J]. Adv ExpMed Biol，1998,440: 675-686.
[117]Pulford D J and Britton P. Intracellular Processing of the Porcine
Coronavirus Transmissible Gastroenteritis Virus Spike Protein Expressed
by Recombinant Vaccinia Virus[J]. Virology. 1991,82(2):765-773.
[118]Wolff J A,Malone R W,Williams P et al. Direct Gene Transfer into Mouse
Muscle in vivo[J]. Science，1990,247:1465-1468.
48 猪传染性胃肠炎病毒 M 基因的克隆及序列分析
[119]Liu C,Kokuho T et al. DNA Mediate Immunization with Encoding the
Nucleoprotein Gene of Porcine Transmissible Gastroenteritis Virus[J].
Virus Res，2001，80(1-2):75-82.
[120]Ulmer J B. Heterologous Protection Against Influenza by Injection of DNA
Encoding a Viral Protein[J]. Science.1993,259(102):1745-1749.
[121]Wang B．Induction of Humoral and Cellular Immune Responses to the Human
Immunodeficiency Type 1 Virus in Nonhuman Primates by in vivo DNA
Inoculation[J].Virology，1995,211(1):102-112.
[122]Kuo L,Godeke G J,Raamsman M J et al. Retargeting of Coronavirus by
Substitution of the Spike Glycoprotein Ectodomain Crossing the Host Cell
Species Barrier[J].Journal of Virology,2000,74:1393－1406.
[123]Spizizen J. Transformation of Biochemically Deficient Strains of Bacillus
Subtilis by Deoxyribonucleate[J]. Proc Natl Acad Sci USA, 1958, 44: 10-22.
[124]Davis H L,Michel M L et al. Direct Gene Transfer into Skeletal
Muscle:Plasmid DNA-based Immunization against the Hepatitis B Virus
Surface Antigen[J].Vaccine，1994,12(16):1503-1509.
[125]Yokoyama M,Zhang J,Whiton J L. DNA immunization Confers Protection against
Lethal Lymphocytic Choriomeningitis Virus Infection[J].J Vrol ，
1995,69:2684-2688.
[126]Xiang Z Q,Spitainilc S L,Cheng J，et al. Immune Responses to Nucleic Acid
Vaccine to Rabies Virus[J]. Vaccine，1995,209:569-579.
[127]Kunst F,Ogasawara N et al. The Complete Genome Sequence of the Grampositive
Model Organism Bacillus subtilis (strain 168)[J].Nature,1997,390:249.
[128]Reiset J. Transfer and Expression of Heterologous Genes in Yeast Other
than Saccharomyces Cerevisrae[J]. Adv Bioch Eng Biotech,1990, 43:75-102.
[129]Austin S,Bingham E T,Koegel R G et al. An Overview of a Feasibility Study
for the Production Enzymes in Transgenic Alfalfa[J]. Ann N Y Acad Sci,
1999,2(721):234-244.
[130]Cox G M,Zamb T J,Babiuk L A. Bovine Herpesvirus 1:Immune Responses in Mice
and Cattle Injected with Plasmid DNA[J]. J Virol. 1993,67(9): 5664-5667.
[131]Sirnarumitr T, Siddell S, Grahare F, et al. Porcine transmissible
gastroenteritis virus induced apoptosis in swine testes cell cultures[J].
Archives of Virology,1998,143(12):2471-2485.
[132]Jones T, Shenk T,Transmissible gastroenteritis virus of pigs[J].
Veterinary R EcoRd,1997,141(16):427-428.
参考文献 49
[133]Woods R D.Development of PCR-based techniques to identify porcine
transmissible gastroenteritis coronavirus isolated[J].Can J Vet
Res,1997,61(3):167-172.
[134]Paton D, Ibata G, Sands J, et al. Detection of transmissible gastroenteritis
virus by RT-PCR and differentiation from porine respiratory
coronavirus[J]. J Virol Methods,1997,66(2):303-309.
[135]Kim O, Chol C, Kim B, et al. Detection and differentiation of porine
epidemic diarrhea virus and transmissible gastroenteritis virus in
clinical sample by multiplex RT-PCR[J]. Vet Res,2000,146(22):637-640.
[136]Eleouet J, Bett A, Koolen M, et al.Complete seqence of the
polyprotein-encoding gene 1 of transmissible gastroenteritis virus[J].
Journal of Virology,1995,206:817-822.
[137]Godet M,Stewart F,Higgins D. Processing and Antigenicity of Entire and
Anchor-free S Protein of TGEV Expressed by Recombinant Baculovirus[J].
Journal of Virol,1991,185:732-740.
[138]Shoup D. Active and passive immune responses to TGEV in Swine Inoculated
with Recombinant Baculovirus-expressed TGEV Spike Glycoprotein
Vaccines[J]. American Journal of Veterinary Research, 1997,58(3):242-250.
研究[J].中国兽医科技,2001,31(8):8-9.
[2] 王继科，刘常明，马思奇,等.猪传染性胃肠炎和猪流行性腹泻病毒免疫电镜的诊断
的研究[J].中国畜禽传染病,1991,2:22-25.
[3] 方定一. 兽医微生物学[M].北京：农业出版社,1979.
[4] 王明俊.兽医生物制品[M].北京：中国农业出版社,1996.
[5] 任晓峰.猪传染性胃肠炎病毒TH-98株S基因的克隆与鉴定.东北农业大学硕士学位
论文[J].2001
[6] 曹军平,朱爱萍,肖作焕,等.猪传染性胃肠炎的诊断与防制[J].中国动物检疫,
2000,6:29.
[7] 唐立杰，李一经.RT-PCR 方法快速诊断猪传染性胃肠炎[J].黑龙江畜牧兽
医,2001,12:26-27.
[8] 蔡宝祥.家畜传染病学[M].北京,中国农业出版社,2001,213-216.
[9] 周仲芳，立力复，罗长保,等.利用重组抗原建立间接酶联免疫吸附试验检测猪传染
性胃肠炎血清抗体[J].中国兽医杂志,2000,26 (10):12-14.
[10] 赵德明，张中秋，沈建中，主译.B. E.斯特劳，S. D.阿莱尔，W. L.蒙加林等,猪
病学[M],中国农业大学出版社,2000,9:263-276.
[11] 殷震，刘景华.动物病毒学[M].北京：科学出版社,1997,671-703.
[12] 冉智光,王玉春.猪传染性胃肠炎病毒分子生物学研究新进展[J].天津畜牧兽
医,1998,4:4-6.
[13] 佟有恩，冯力，李伟杰，等. 猪传染性胃肠炎与猪流行性腹泻二联弱毒疫苗的研
究[J]. 中国预防兽医学报，1999,21(6):406-410
[14] 师东方.猪轮状病毒和传染性胃肠炎病毒核酸免疫的研究[J]. 东北农业大学博士
学位论文. 2002.
[15] 孙树汉. 核酸疫苗[M]. 上海：第二军医大学出版社，1999.
[16] 梁雨等. 核酸免疫可特异性地诱发小鼠对乙型肝炎病毒表面抗原 S 的免疫反应
[J]。病毒学报，1996,12(3):193.
[17] 王军阳. 核酸疫苗的安全性及伦理学问题[J]。中国医学伦理学，2000,2: 54.
[18] 萨姆布鲁克 J, 弗里奇 E F, 曼尼阿蒂斯 T. 分子克隆实验指南[M]. 金冬雁，
黎孟枫，等. 第二版. 北京:科学出版社,1993,26,674-683.
[19] 邹勇, 钱永清. 猪流行性腹泻、猪传染性胃肠炎和猪轮状病毒 RT-PCR 鉴别诊断技
术研究[J]. 上海农业学报,2003,19(2):82-84.
[20] 吴凌, 李一经. 应用 RT-nested PCR 检测猪流行性腹泻病毒的研究[J]. 中国兽医
40 猪传染性胃肠炎病毒 M 基因的克隆及序列分析
科技,2003,33(3):27-30.
[21]乔宪凤，熊忠良，郑新民等.PEDV 膜蛋白基因 PT-PCR 最适反应条件的确立[J].湖北
畜牧兽医，2001，1，8-10.
[22] Saif L J,Wesley R D.Transmissible gastroenteritis In Diseases of Swine[M].
Iowa State University Press,Ames,IA,1992,362-386.
[23] Enjuanes L,Van der Zeijst B A M. Molecular Basis of Transmissible
Gastroenteritis Coronavirus Epidemiology[M].PlenumPress,New York,N.Y.
1995, 337-376.
[24] Paton D, Ibata G, Sands J, McGoldrick A.Detection of Transmissible
Gastroenteritis Virus by RT-PCR and Differentiation from Porcine
Respiratory Coronavirus[J].J Virol Methods,1997,66(2):303-309.
[25] Enjuanes L, Spaan W, Snijder E Cavanagh D. Nidovirales. Classification and
Nomenclature of Viruses In Virus taxonomy[M].New York,Academic Press,2000,
827-834.
[26] Wesley RD,Woods R and Cheuny A. Genetic Basis of the Pathogenesis of
Transmissible Gastroenteritis Virus[J].Journal of Virol,1990,64(10):
4761-4766.
[27] Paton D, Lowings P. Discrimination between Transmissible Gastroenteritis
virus Isolates[J].Arch Virol.1997b,142(8):1703-1711.
[28] Goodwin R F W ,Jennings A R.A Highly Infectious Gastroenteritis of Pig[J].
Vet Rec,1958,70:271-272.
[29] Sasahara J, Harada K, Hayashi S and Watanabe M.Studies on Transmissible
Gastroenteritis in Pig in Japan. Japan[J]. J. Vet.Sci,1958,20:1-6.
[30] Siddell S G. The Coronavifdae In The Viruses[M], New York,Plenum Press.
1995,418.
[31] Lai M C ,Cavanagh D. The Molecular Biology of Coronaviruses[J]. Advances
in Virus Research,1997,48:1-100.
[32] Caul E ,Egglestone S.Coronavirus in Humans In Virus Infections of the
Gastrointestinal Tract[M]. New York: Marcel Dekker,1982,179-193.
[33] Denison M R. In Viral Infections of the Respiratory Tract Edited by
Dolin[M]. New York,Marcel Dekker,1999,253-280.
[34] Enjuanes L, Sola I, Almazan F, Ortego J, Izeta A, Gonzalez J M, Alonso S,.
Coronavirus Derived Expression Systems[J].Journal of Biotechnology,2001，
88:183-204.
[35] Kemeny L J.Antibody Response in Pigs Inoculated with Transmissible
参考文献 41
Gastroenteritis Virus and Cross Reactions among Ten Isolates[J].Can J Comp
Med.1976,40: 209-214.
[36] Pederden M C, Ward J and Mengeling W L.Antigenic Relationship of Feline
Infectious Peritonitis Virus to Coronaviruses of Other Species[J].Arch.
Virol,1978,58:45-53.
[37] Reynolds D J, Garwes D J and Lucey S.Differentiation of Canine Coronavirus
and Porcine Transmissible Gastroenteritis Virus by Neutralization with
Canine，Porcine and Feline Sera[J].Vet. Microbiol,1980,5:283-290.
[38] Woods R D.Studies of Enteric Coronaviruses in a Feline Cell Line[J].Vet.
Microbiol,1982,7:427-435.
[39] Haeltennan E O.Epidemiological Studies of Transmissible Gastroenteritis
of Swine[J].Proc US Livest Sunit Assoc,1962,66:305-315.
[40] Klemm R C,Ristic M.The Effect of Propagation of Transmissible
Gastroenteritis(TGE) Virus in Pups and the Lung of Baby Pigs on the
Immunologic Properties of the VirusInt Congr Pig Vet Soc[M].Iowa State
Univ,1976,11.
[41] Pensaert M, Callebaut P and Vergote J.Isolation of a Porcine Respiratory,
Non-enteric Coronavirus Related to Transmissible Gastroenteritis[J].Vet
Q. 1986,8:257-261.
[42] Kim B, Kim O, Tai J H, Chae C.Transmissible Gastroenteritis Virus Induces
Apoptosis in Swine Testicular Cell Lines But not in Intestinal
Enterocytes[J].J Com Pathol,2000,123(1):64-66.
[43] Hooper B E and Haelterman E O.Concepts of Pathogenesis and Passive Immunity
in Transmissible Gastroenteritis of Swine[J].J Am Vet Med Assoc,
1966,149:1580-1586.
[44] Butler D G, Gall D G, Kelly M H,et al.Transmissible Gastroenteritis
Mechanisms Responsible for Diarrhea in an Acute Enteritis in Piglets[J].
J Clin Invest,1974,53:1335-1342.
[45] Comelius L M, Hooper B E and Haelterman E O. Changes in Fluid and Electrolyte
Balance in Baby Pigs with Transmissible Gastroenteritis[J].Am J Clin Pathol,
1968,2:105-113.
[46] Wagner J E, Beamer P D and Ristic M.Electron Microscopy of Intestinal
Epithelial Cells of Piglets Infected with a Transmissible Gastroenteritis
Virus[J].Can J Comp Med,1973,37:177-188.
[47] Dul G C, Ruckerbauer P,Transmissible Gastroenteritis Demonstration of the
42 猪传染性胃肠炎病毒 M 基因的克隆及序列分析
Virus from Field Specimens by Means of Cell Culture and Pig Inoculation[J].
Can J Comp Med,1977,41:357-363.
[48] Bohl E H. Diagnosis of Diarrhea in Pigs due to Transmissible Gastroenteritis
Virus or Rotavirus In Viral Enteritis in Humans and Animals[J].Ed Bricout
and Scherrer INSERM,1979,90:341-343.
[49] Stark S L, Fernelius A L, Booth G D et al.Transmissible gastroenteritis
(TGE) of swine Effect of Age of Swine Testes Cell Culture Monolayers on Plaque
Assays of TGE Virus[J].Can J Comp Med,1975,39:466-468.
[50] Black J W.Diagnosis of TGE by FA Evaluation of Accuracy on Field
Specimens[J].Proc US Anim Health Assoc,1971,75:492-498.
[51] Garwes D J, Stewart F, Cartwright S F et al. Differentiation of Porcine
Coronavirus from Transmissible Gastroenteritis Virus. Vet Rec. 1988,122:
86-87.
[52] Van Nieuwstadt A P and Pol J M A. Isolation of a TGE Virus-related Respiratory
Coronavirus Causing Fatal Pneumonia in Pigs[J].Vet Rec,1989,124:43-44.
[53] Benfield D A, Jackwood D J, Bae I,et al.Detection of Transmissible
Gastroenteritis Virus Using cDNA Probes[J].Arch Virol,1991,116:91-106.
[54] Paton D,Lowings P.Discrimination between Transmissible Gastroenteritis
virus Isolates[J].Arch Virol,1997,142(8):1703-1711.
[55] Van Cott J L, Brim T A, Simkins R A,et al.Antibody-secreting Cells to
Transmissible Gastroenteritis Virus and Porcine Respiratory Coronavirus
in Gut and Bronchus-associated Lymphoid Tissues of Neonatal Pigs[J].J
Immunol, 1993,150:3990-4000.
[56] Saif L J, Van Cott J L ,Brim T A.Immunity to Transmissible Gastroenteritis
Virus and Porcine Respiratory Coronavirus Infections in Swine[J].Vet
Immunol Immunopathol,1994,43:89-97.
[57] Anton I M, Gonzalez S, Bullido M J,et al.Cooperation Between Transmissible
Gastroenteritis Coronavirus (TGEV) Structural Proteins in the in vitro
Induction of Virus-specific Antibodies[J].Virus Res,1996,46:111-124.
[58] Siddle S G, Anderson R, Cavanagh D, Fujiwara K, Coronaviridae[J].
Intervirology,1983,20:181-189.
[59] Spaan W,Cavanagh D,Horizinek M C.Coronaviruses Structure and Gene
Expression[J].Journal of Gen Virol,1988,69:2939-2952.
[60] Britton P and Page D W. Sequence of the S Gene from a Virulent British Field
Isolate of Transmissible Gastroenteritis Virus[J].Journal of Virus Res,
参考文献 43
1990,18:71-80.
[61] Curtis K M, Yount B,Baric R S.Heterologous Gene Expression from
Transmissible Gastroenteritis Virus Replicon Particles[J]. Journal of
Virol,2002,76(3):1422-1434.
[62] Cavanagh D, Brian D A, Brinton M A et al. Revision of the Taxonomy of the
Coronaviruses Torovirus and Arterivirus Genera[J].Archive of Virol,1994,
135(2):227-237.
[63] Kevin W, Paul P, Britton ,Michael E G. Sequence Analysis of the Leader RNA
of Transmissible Gastroenteritis Virus[J].Virus Gene,1990,4:289-303.
[64] Hiscox J A.Characterization of the Transmissible Gastroenteritis Vinis
(TGEV) Transcription Initiation Sequence[J].Adv Exp Med Biol,1995,
380:529-535.
[65] Wesley R, Woods R and Cheuny, A. Nucleotide Sequence of the E2-peplomer
Protein Gene and Partial Nucleotide Sequence of Upstream Polymerase Gene
of Transmissible Gastroenteritis Virus[J].Adv Exp Med Biol,
1990,276:301-306.
[66] Paul B, Kevin W. The Cloning and Sequencing of the Virion Protein Genes
from a British Isolate of PRCV Comparison with TGEV[J]. Journal of Virus
Research, 1991,21:181-198.
[67] Julian A, Hiscox, David C et al.Quantification of Individual Subgenomic
mRNA Species during Replication of the Transmissible Gastroenteritis
Virus[J]. Virus Res,1995,36:119-130.
[68] Gebauer F, Posthumus W P, Correa 1, Sune C, Smerdou C.Residues Involved
in the Antigenic Sites of Transmissible Gastroenteritis Coronavirus S
Glycoprotein[J].Virology,1991,83(1):225-238.
[69] Chen C M, Cavanagh D,Britton P.Cloning and Sequencing of a 8.4kb Region
from the 3'End of a Taiwanese Virulent Isolate of the Coronavirus
Transmissible Gastroenteritis Virus[J].Virus Res,1995,38(1):83-89
[70] Ballesteros L, Sanchez C ,Enjuanes L.Two Amino Acid Changes at the
N-terminus of TGEV Spike Protein Result in the Loss of Enteric
Tropism[J].Journal of Virol,1997,227(2):378-388.
[71] Christine K, Graham D, Yolken R et al.Point Mutations in the S Protein
Connect the Sialic Acid Binding Activity with the Enteropathogenicity of
TGEV[J]. Journal of Virol,1997,71(4):3285-3287.
[72] Krempl C, Schultze B, Laude H,Healer G.Point Mutations in the S Protein
44 猪传染性胃肠炎病毒 M 基因的克隆及序列分析
Connect the Sialic Acid Binding Activity with Enteropathogenicity of
Transmissibl Gastroenteritis
Virus[J]JournalofVirol,1997,71(4):3285-3287
[73] Delmas B, Rasschaert D, Gtodet M, Gelfi J.Four Major Antigenic Site of the
Coronavirus Transmissible Gastroenteritis Virus are Located on the
Amino-terminal Half of Spike Glycoprotein S[J].Journal of Gen Virol,
1990,71:1313-1323.
[74] Laude H, Rasschaert D, Delmas B,et al.Molecular Biology of Transmissible
Gastroenteritis Virus[J].Vet Virol,1990,23:147-154.
[75] Gebauer F, Posthumus W P, Correa 1, Sune C, Smerdou C.Residues Involved
in the Antigenic Sites of Transmissible Gastroenteritis Coronavirus S
Glycoprotein[J].Virology,199183(1):225-238.
[76] Correa I, Jimenez G, Sune C, Bullido M J.Antigenic Structure of the E2
Glycoprotein from Transmissible Gastroenteritis Coronavirus[J].Virus Res,
1988,10:77-94.
[77] Tuboly T, Nagy E.Immunogenicity of the S Protein of Transmissible
Gastroenteritis Virus Expressed in Baculo virus[J].Arch Virol,
1994,137(2):55-67.
[78] Bernard S and Laude H.Site-specific Alteration of Transmissible
Gastroenteritis Virus Spike Protein Results in Markedly Reduced
Pathogenicity. Journal of Gen[J].Virol,1995,76:2235-2241.
[79] Christine K, Graham D, Yolken R,et al.Point Mutations in the S Protein
Connect the Sialic Acid Binding Activity with the Enteropathogenicity of
TGEV[J].Journal of Virol,1997,71(4):3285-3287.
[80] Laviada M D, Videgain S P, Moreno L M.Expression of Swine Transmissible
Gastroenteritis Virus Envelope Antigens on the Surface of Infected Cell:
Epitopes externally exposed[J].Virus Res,1990,16:247-254.
[81] Sune C, Jimenez G, Correa I.Mechanisms of Transmissible Gastroenteritis
Virus Coronavirus Neutralization[J].Virology,1990,177:559-569.
[82] Delmas B, Rasschaert D, Gtodet M.Four Major Antigenic Site of the
Coronavirus Transmissible Gastroenteritis Virus are Located on the
Amino-terminal Half of Spike Glycoprotein S[J].Journal of Gen
Virol,1990,71:1313-1323.
[83] Delmas B, Gelfi J.Antigenic Structure of Transmissible Gastroenteritis
Virus Domains in the Peplomer Glycoprotein[H].Journal of Virol,1986,67:
参考文献 45
1405-1418.
[84] Have P.Infection with a New Porcine Respiratory Coronavirus in Denmark:
Serologic Differentiation from Transmissible Gastroenteritis Virus Using
Monoclonal Antibodies[M].In Coronaviruses and Their Diseases Plenum Press,
New York,1990,435-439.
[85] Sanchez C, Jimenez G, Laviada M D.Antigenic Homology among Coronavirus
Related to Transmissible Gastroenteriti Virus[J].Virology,1990,174:414
[86] Kim L, Hayes J. Molecular Characterization and Pathogenesis of
Transmissible Gastroenteritis Coronavirus (TGEV) and Respiratory
CoronavirusRC Field IsolatesCo-circulatingina Swine Herd[J].Arch
Virology,2000,145:1133-1147.
[87] Simpkins R A, Weilnau P A.Antigenic Variation among Transmissible
Gastroenteritis Virus (TGEV) and Porcine Respiratory Coronavirus Strains
Detected with Monoclonal Antibodies to the S Protein of TGEV[J].Am. J.
Vet. Res,1992,53:1253-1258.
[88] Niemann H, Boschek B, Evans D.Post-translational Glycosylation of
Coronavirus Glycoprotein E1 Inhibition by Monensin[J].Eur Mol Biol
Organ,1982,1:1499-1504.
[89] Jimenez G, Correa I, Melgosa M P.Critical Epitopes in Transmissible
Gastroenteritis Virus Neutralization[J].Journal of Virol,1986,60:131
一 139.
[90] Jones T ，Shenk T.Transmissible Gastroenteritis Virus of pigs[J].Vet Rec,
1997,41(16):427-428.
[91] Jacobs L, Van Der Zeust B A M,Horzined M C. Characterization and Translation
of Transmissible Gastroenteritis Virus RNAs[J].Journal of Virol,1986,57:
1010-1015.
[92] Carlos M. Sanchez, Izeta A, Jose M. Targeted Recombination Demonstrates
that the Spike Gene of Transmissible Gastroenteritis Coronavirus is a
Determinant of Its Enteric Tropism and Virulence[J].Journal of
Virol,1999,73:7607-7618.
[93] Taguchi F ，Fleming J O. Comparison of Six Different Murine Coronavius JHM
Variants by Monoclonal Antibodies against the E2 Glycoprotein[J].
Virology,1989,169:223-235
[94] Collins A R, Knobler R L, Powell H .Monoclonal Antibodies to Murine Hepatitis
Virus-4(strain JHM)Define the Viral Glycoprotein Responsible for
46 猪传染性胃肠炎病毒 M 基因的克隆及序列分析
Attachment and Cell-cell Fusion[J].Virology,1982,119:358-371.
[95] Sturman L S, Richard C S,Murine K V. Proteolytic Cleavage of the E2
Glycoprotein of Murine Coronavirus by Trysin and Separation of Two
Different 90K Cleavage Fragments[J].Journal of Virol,1985,56:904-911.
[96] Ballesteros L, Sanchez C ,Enjuanes L.Two Amino Acid Changes at the
N-terminus of TGEV Spike Protein Result in the Loss of Enteric
Tropism[J].Journal of Virol,1997,227(2):378-388.
[97] Christine K, Graham D, Yolken R et al. Point Mutations in the S Protein
Connect the Sialic Acid Binding Activity with the Enteropathogenicity of
TGEV Journal of Virol. 1997,71(4):3285-3287.
[98] Motokawa K. Comparison of the Amino Acid Sequence and Phylogenetic Analysis
of the Peplomer Integral Membrane and Nucleocapsid Protein of Feline Canine
Porcine Corona virus[J].Journal of Microbiology and immunology.
1996,40(6):425-433.
[99]Rasschaert D ,Laude H. The Predicted Primary Structure of the Peplomer
Protein E2 of the Porcine Coronavirus Transmissible Gastroenteritis
Virus[J].Journal of Gen Virol,1987,68:1883-1890.
[100]Charley B ， Laude H. Induction of Alpha Interferon by Transmissible
Gastroenteritis is Coronavirus:Role of Transmembrane Glycoprotein
E1[J].Journal of Virol,1988,62:8-11.
[101]Laude H, Rasschaert D, Delmas B,et al.Molecular Biology of Transmissible
Gastroenteritis Virus[J].Vet Virol,1990,23:147-154.
[102]Garwes D, Pocock D H,Wllaszka T M.The Polypeptide Structure of
Transmissible Gastroenteritis Virus[J].Journal of Gen Vrol,1975,29:25-34.
[103]Motokawa K.Comparison of the Amino Acid Sequence and Phylogenetic Analysis
of the Peplomer Integral Membrane and Nucleocapsid Protein of Feline Canine
and Porcine Coronavirus[J].Journal of Microbiology and Immunology,
1996,40(6):425-433.
[104]Nakange K,Yamanouchi K,Fujiwara K.Protective Effect of Monoclonal
Antibodies on Lethal Mouse Hepatitis Infection in Mice[J].Journal of Virol,
1986,59:168-171.
[105]Compton J，Rogers D B, Holmes K V, Fersch D.In vitro Replication of Mouse
Hepatitis Virus Strain A59[J].Journal of Virol,1987,61:1814-1820.
[106]Baric R S, Nelson G W J O, Deans R J.Interaction Between Coronavirus
Nucleocapsid Protein and Viral RNA Implications for Viral Transcription[J].
参考文献 47
Journal of Virol,1988,62:4280-4287.
[107]Stohlman L M, Baric R, Nelson G N, Welter L M .Specific Interaction between
Coronavirus Leader RNA and Nucleocapsid Protein[J].Journal of Virol,
1988,62:4288-4295.
[108]Boursnell M E G, Brown T D K, Foulds I J, Green P F. Completion of the Sequence
of the Genome of the Coronavirus Avian Infectious Bronchitis Virus[J].
Journal of Gen Virol,1987,68: 53-77.
[109]Wesley R D,Woods R,Cheuny A.Genetic Basis of the Pathogenesis of
Transmissible Gastroenteritis Virus[J]. Journal of Virol,1990,64(10):
4761-4766.
[110]Wesley R D, Woods R D ,Cheung A K.Genetic Analysis of Porcine Respiratory
Coronavirus an Attenuated Variant of Transmissible Gastroenteritis
Coronaviurs[J].Journal of Virol,1991,65:3369-3373.
[111]Julian A, Hiscox, David C ,et al.Quantification of Individual Subgenomic
mRNA Species during Replication of the Transmissible Gastroenteritis
Virus[J].Virus Res,1995,36:119-130.
[112]Leman A D, Straw B, Glock R D,et al.Disease of Swine[M] .Ames Iowa state
university press,1992:362-386.
[113] Garwes D J,Lucas M H,Higging D A,Pike B V ，Cratwright S F. Antigenicity
of Structural Components from Porcine Transmissible Gastroenteritis
Virus[J]. Vet Microbiol，1978,1979,3:179-190.
[114]Gough P M,Frank C J,Moore D G，et al. Lactogenic Immunity to Transmissible
Gastroenteritis Virus Induced by a Subunit Immunogen[J].Vaccine ，
1983,1:37-41.
[115]Posthumus W P,Enstra J A, Schaaper W M ，et al. Analysis and Simulation
of Neutralizing Epitope of Transmissible Gastroenteritis Virus[J]. J
Virol，1990,64:3304-3309.
[116]Castilla J,Sola I,Pintado B,Sanchez-Morgado J M，Enjuanes L. Lactogenic
Immunity in Transgenic Mice Producing Recombinant Antibodies Neutralizing
Coronavirus[J]. Adv ExpMed Biol，1998,440: 675-686.
[117]Pulford D J and Britton P. Intracellular Processing of the Porcine
Coronavirus Transmissible Gastroenteritis Virus Spike Protein Expressed
by Recombinant Vaccinia Virus[J]. Virology. 1991,82(2):765-773.
[118]Wolff J A,Malone R W,Williams P et al. Direct Gene Transfer into Mouse
Muscle in vivo[J]. Science，1990,247:1465-1468.
48 猪传染性胃肠炎病毒 M 基因的克隆及序列分析
[119]Liu C,Kokuho T et al. DNA Mediate Immunization with Encoding the
Nucleoprotein Gene of Porcine Transmissible Gastroenteritis Virus[J].
Virus Res，2001，80(1-2):75-82.
[120]Ulmer J B. Heterologous Protection Against Influenza by Injection of DNA
Encoding a Viral Protein[J]. Science.1993,259(102):1745-1749.
[121]Wang B．Induction of Humoral and Cellular Immune Responses to the Human
Immunodeficiency Type 1 Virus in Nonhuman Primates by in vivo DNA
Inoculation[J].Virology，1995,211(1):102-112.
[122]Kuo L,Godeke G J,Raamsman M J et al. Retargeting of Coronavirus by
Substitution of the Spike Glycoprotein Ectodomain Crossing the Host Cell
Species Barrier[J].Journal of Virology,2000,74:1393－1406.
[123]Spizizen J. Transformation of Biochemically Deficient Strains of Bacillus
Subtilis by Deoxyribonucleate[J]. Proc Natl Acad Sci USA, 1958, 44: 10-22.
[124]Davis H L,Michel M L et al. Direct Gene Transfer into Skeletal
Muscle:Plasmid DNA-based Immunization against the Hepatitis B Virus
Surface Antigen[J].Vaccine，1994,12(16):1503-1509.
[125]Yokoyama M,Zhang J,Whiton J L. DNA immunization Confers Protection against
Lethal Lymphocytic Choriomeningitis Virus Infection[J].J Vrol ，
1995,69:2684-2688.
[126]Xiang Z Q,Spitainilc S L,Cheng J，et al. Immune Responses to Nucleic Acid
Vaccine to Rabies Virus[J]. Vaccine，1995,209:569-579.
[127]Kunst F,Ogasawara N et al. The Complete Genome Sequence of the Grampositive
Model Organism Bacillus subtilis (strain 168)[J].Nature,1997,390:249.
[128]Reiset J. Transfer and Expression of Heterologous Genes in Yeast Other
than Saccharomyces Cerevisrae[J]. Adv Bioch Eng Biotech,1990, 43:75-102.
[129]Austin S,Bingham E T,Koegel R G et al. An Overview of a Feasibility Study
for the Production Enzymes in Transgenic Alfalfa[J]. Ann N Y Acad Sci,
1999,2(721):234-244.
[130]Cox G M,Zamb T J,Babiuk L A. Bovine Herpesvirus 1:Immune Responses in Mice
and Cattle Injected with Plasmid DNA[J]. J Virol. 1993,67(9): 5664-5667.
[131]Sirnarumitr T, Siddell S, Grahare F, et al. Porcine transmissible
gastroenteritis virus induced apoptosis in swine testes cell cultures[J].
Archives of Virology,1998,143(12):2471-2485.
[132]Jones T, Shenk T,Transmissible gastroenteritis virus of pigs[J].
Veterinary R EcoRd,1997,141(16):427-428.
参考文献 49
[133]Woods R D.Development of PCR-based techniques to identify porcine
transmissible gastroenteritis coronavirus isolated[J].Can J Vet
Res,1997,61(3):167-172.
[134]Paton D, Ibata G, Sands J, et al. Detection of transmissible gastroenteritis
virus by RT-PCR and differentiation from porine respiratory
coronavirus[J]. J Virol Methods,1997,66(2):303-309.
[135]Kim O, Chol C, Kim B, et al. Detection and differentiation of porine
epidemic diarrhea virus and transmissible gastroenteritis virus in
clinical sample by multiplex RT-PCR[J]. Vet Res,2000,146(22):637-640.
[136]Eleouet J, Bett A, Koolen M, et al.Complete seqence of the
polyprotein-encoding gene 1 of transmissible gastroenteritis virus[J].
Journal of Virology,1995,206:817-822.
[137]Godet M,Stewart F,Higgins D. Processing and Antigenicity of Entire and
Anchor-free S Protein of TGEV Expressed by Recombinant Baculovirus[J].
Journal of Virol,1991,185:732-740.
[138]Shoup D. Active and passive immune responses to TGEV in Swine Inoculated
with Recombinant Baculovirus-expressed TGEV Spike Glycoprotein
Vaccines[J]. American Journal of Veterinary Research, 1997,58(3):242-250.