牛传染性鼻气管炎TK/gE基因缺失重组病毒的研究
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摘要
牛传染性鼻气管炎(Infectious Bovine Rhinotracheitis, IBR),是由牛疱疹病毒1型(BHV-1)引起的一种牛的急性、热性、接触性传染病,主要引起呼吸道和生殖道疾病,如传染性鼻气管炎、眼结膜炎、高热、传染性脓疱性外阴道炎、流产等。自被发现以来,该病就在世界范围内流行,给养牛业造成巨大的经济损失。IBR被国际兽疫局(0IE)列为B类疾病,该病的根治方法是阳性牛检出与捕杀,而防制的主要措施是接种疫苗,主要采用基因工程疫苗的方法,国外也已相继开发出了牛疱疹病毒1型TK、gE基因缺失重组疫苗。
BHV-1是有囊膜的双股DNA病毒,其基因组G+C含量为72%,大小约为138kb,由一个UL区(106kb)和一个US区(10kb)及US区两侧的重复序列(IRs和TRs,各1lkb,序列相同但方向相反)组成。BHV-1基因组编码70个左右基因,其中有10种为糖蛋白。其中gE糖蛋白是病毒囊膜上的主要糖蛋白。它以细胞特异的方式影响病毒从被感染细胞中释放,同时gE基因的缺失也影响到病毒在细胞间的传播。而胸腺激酶(TK)基因是牛疱疹病毒1型增殖的非必需基因,也是主要毒力基因之一,对病毒在神经组织的持续感染具有非常重要的作用。
本研究构建了牛疱疹病毒1型TK／gE双基因缺失突变株,并做了动物安全性和保护力试验,为研究缺失标志活病毒疫苗及研制表达外源基因的BHV-1重组病毒活载体疫苗奠定基础。主要工作包括：1.牛传染性鼻气管炎TK~-／gE~-基因双缺失突变株的构建
1)牛传染性鼻气管炎TK-/EGFP+突变株的构建
本实验以牛疱疹病毒1型全序列(基因登录号为AJ004801)为基础,提取病毒全基因组。在此基础上设计引物,用PCR技术扩增TK基因上游和下游约1.0 kb和1.1kb的片段作为同源重组臂,分别克隆于载体pBluescriptⅡSK(+)中。用相应的限制性内切酶将下游同源臂回收,定向插入到上游的克隆载体中,构建中间转移载体,并命名为pZF08-21。然后插入带有完整启动子的EGFP报告基因,构建成重组转移质粒,并命名为pZF07-16。以磷酸钙转染法将经线性化的pZF07-16、质粒pBICP0与BHV-1基因组DNA在牛肾细胞(MDBK)上共转染,成功地进行了同源重组,7轮筛选和纯化获得了BHV-1 TK-/EGFP+突变株。
2)牛传染性鼻气管炎TK~-突变株的构建
提取BHV-1 TK-/EGFP+突变株基因组,再次以磷酸钙转染法将经线性化的pZF08-21、质粒pBICP0与基因组DNA在MDBK上共转染,同源重组去掉EGFP荧光标签,经5轮纯化成功的获得了BHV-1 TK-突变株。
3)牛传染性鼻气管炎TK-/gE-/EGFP+双缺失突变株的构建
方法同1,用PCR扩增gE基因上游和下游约1.0kb和1.1kb的片段,分别克隆于载体pcDNA3.1(+)myc-His B中。通过限制性酶切将上游同源臂回收,插入到下游的克隆载体中,构建中间转移载体,并命名为pZF09-08。然后插入EGFP报告基因,构建成重组转移质粒,并命名为pZF09-15。以磷酸钙转染法将经线性化的pZF09-15、质粒pBICP0与BHV-1 TK"缺失突变株基因组DNA在MDBK细胞上共转染,成功地进行同源重组,经过7轮筛选和纯化获得了BHV-1 TK-/gE-/EGFP+突变株。
2.牛传染性鼻气管炎TK-/gE-/EGFP+双缺失突变株的安全性和保护力研究
1)牛传染性鼻气管炎TK-/gE-/EGFP+双缺失突变株对牛的安全性研究
安全性研究包括三部分：接种试验、同居试验、再激活试验。接种试验和同居试验：将试验组按剂量分为105,106,107三栏,滴鼻免疫4X105、4X106、4X107PFU的双缺失突变株,另设空白对照组。另在各栏中同时喂养未接种的牛作为同居对照。测量试验牛的体温,观察精神状态和采食情况,采集检测样品。再激活试验：接种免疫28天后,连续3天以15mg／头的剂量肌肉注射地塞米松,以激活处于潜伏感染状态的病毒。试验证实了牛传染性鼻气管炎TK-/gE-/EGFP+双缺失突变株具有很高的安全性。
2)牛传染性鼻气管炎TK-/gE-/EGFP+双缺失突变株对牛的保护力研究
两月龄牛分别用4X105、4X106、4X107PFU的双缺失突变株免疫4周后,以4X107PFU BHV-1强毒攻击。同时设免疫DMEM的空白对照组。分别于攻毒后1d、2d、3d、5d、7d、14d、21d和28d测量体温、采集检测样品。试验结果表明：牛传染性鼻气管炎TK-/gE-/EGFP+双缺失突变株对牛具有很强的保护力。
总之,本研究表明,BHV-1 TK-/gE-可以作为候选疫苗毒株,为我国牛传染性鼻气管炎根除计划提供工具。
Infectious bovine rhinotracheitis (IBR), caused by bovine herpesvirus type 1 (BHV-1), is known as an acute, pyrexic and contagious disease. It's a major pathogen of cattle causing respiratory and genital tract infections such as bovine rhinotracheitis, conjunctivitis, fever, infectious pustular vulvovaginitis, and abortion. Since BHV-1 was discovered, IBR has occurred all over the world, and causes great economic loss to the cattle and milk industry. IBR is ranked in list B diseases by OIE, and the main measure for eradication is to slaughter the infected cattle. In addition to this, Immunization is an effective way to control it. Many live BHV-1 vaccines has been developed such as BHV-1 TK null, BHV-1 gE null and so on.
The genome of BHV-1 is a double-strand DNA of about 138 kb, with the G+C content of 72%. It contains two unique areas called the long unique (UL) and the short unique(US), and two reverse repeat units, e.t. IR and TR, flanking the US area. BHV-1 encodes 10 glycoproteins, including glycoprotein E (gE). gE is a non-essential gene for virus growth, but it influences cell-to-cell transmission in MDBK cell monolayer. On the other hand, thymidine kinase (TK) gene, another main virulence determing gene, is also non-essential for virus proliferation. It plays a very important role in the latency infection in the nervous system.
The objective of this study is to construct a bovine herpersvirus type 1 live modified vaccine strain with TK gene and gE gene deletion, providing a platform for a live viral vector expressing exogenous genes. Safety and efficacy of the both TK gene and gE gene deleted vaccine strain was carried out. The research results are summarized as follows:
1. Construction of BHV-1 TK-/gE-/EGFP+ double gene-deleted mutants
1). Construction of BHV-1 TK-/EGFP+ gene-deleted mutant
Two pairs of primers of TK gene was synthesized based on the genomic DNA sequence of BHV-1 (GenBank Accession No. AJ004801). The upstream (1.0kb) and downstream (1.1kb) of TK gene homologous arms were amplified by PCR, sequenced, and were directly cloned into pBluescriptⅡSK(+) vector one by one by restriction enzymes. Then EGFP expression cassette was inserted in between the two homologous arms. The recombinant plasmid was linearized and co-transfect into MDBK cells together with pBICPO and BHV-1 wild-type genomic DNA. By green fluorescence screening and plaque purification, the recombinant BHV-1 TK-/EGFP+ was generated.
2). Construction of BHV-1 TK- gene-deleted mutant
The pZF08-21 was linearized and co-transfected into MDBK cells together with pBICPO and BHV-1 TK-/EGFP+ genomic DNA. Through five rounds of flurescence selection and plaque purification, EGFP expression cassette was successfully removed, and BHV-1 TK- gene-deleted virus was generated.
3). Construction of BHV-1 TK-/gE-/EGFP+ double gene-deleted mutant
Two pairs of primers of gE gene was synthesized based on the genomic DNA sequence of BHV-1 (GenBank accession No. AJ004801). The upstream (1.1kb) and downstream (1.0kb) of gE gene homologous arms were amplified by PCR, sequenced, and were directly cloned into pcDNA3.1(+) myc-His B vector one by one by restriction enzymes. The EGFP expression cassette was inserted in between the two homologous arms. The transfer plasmid was linearized and co-transfected into MDBK cells together with pBICPO and BHV-1 TK- genomic DNA. Through nine rounds of flurescence selection and plaque purification, BHV-1 TK-/gE-/EGFP+ double gene-deleted mutant was generated.
2. The safety and efficacy of BHV-1 TK-/gE-/EGFP+ double gene-deleted vaccine strain in cattle
1)The safety of BHV-1 TK-/gE-/EGFP+ double gene-deleted mutant
The safety experiment was divided into three parts.They were inoculation experiment, cohabitation experiment and reactivation experiment. Three different immune dose of vaccine strain (4×105PFU,4×106PFU,4×107PFU) were infected intranasally in 10 cattle. In addition, unvaccinated cattle was designated as cohabitation test in the inoculation group. Body temperature was mesured daily, serum samples and swabs were collected. Then, dexamethasone were injected for three days to reactivate the latent virus.The safe experiment shows that the TK-/gE-/EGFP+ gene-deleted mutant is quite safe for cattle.
2)The efficacy of BHV-1 TK-/gE-/EGFP+ double gene-deleted mutant
Vaccination group and negative control group were classified in this experiment. Three different immune dose of vaccine strain (4×105PFU,4×106PFU,4×107PFU) were used to infect 9 cattle of two month old in the vaccination group, while DMEM were used in the negative control group (6 cattle). Four weeks post inoculation, both vaccination group and the control group were challenged with 4×107PFU BHV-1 wild-type virus intranasally. Body temperature was mesured daily, serum samples and swabs were collected on 1d,2d, 3d,5d,7d,14d,21d and 28d post challenging. The efficacy experiment shows that the BHV-1 TK-/gE- double gene-deleted virus could provide full immunoprotectivity to cattle.
BHV-1是有囊膜的双股DNA病毒,其基因组G+C含量为72%,大小约为138kb,由一个UL区(106kb)和一个US区(10kb)及US区两侧的重复序列(IRs和TRs,各1lkb,序列相同但方向相反)组成。BHV-1基因组编码70个左右基因,其中有10种为糖蛋白。其中gE糖蛋白是病毒囊膜上的主要糖蛋白。它以细胞特异的方式影响病毒从被感染细胞中释放,同时gE基因的缺失也影响到病毒在细胞间的传播。而胸腺激酶(TK)基因是牛疱疹病毒1型增殖的非必需基因,也是主要毒力基因之一,对病毒在神经组织的持续感染具有非常重要的作用。
本研究构建了牛疱疹病毒1型TK／gE双基因缺失突变株,并做了动物安全性和保护力试验,为研究缺失标志活病毒疫苗及研制表达外源基因的BHV-1重组病毒活载体疫苗奠定基础。主要工作包括：1.牛传染性鼻气管炎TK~-／gE~-基因双缺失突变株的构建
1)牛传染性鼻气管炎TK-/EGFP+突变株的构建
本实验以牛疱疹病毒1型全序列(基因登录号为AJ004801)为基础,提取病毒全基因组。在此基础上设计引物,用PCR技术扩增TK基因上游和下游约1.0 kb和1.1kb的片段作为同源重组臂,分别克隆于载体pBluescriptⅡSK(+)中。用相应的限制性内切酶将下游同源臂回收,定向插入到上游的克隆载体中,构建中间转移载体,并命名为pZF08-21。然后插入带有完整启动子的EGFP报告基因,构建成重组转移质粒,并命名为pZF07-16。以磷酸钙转染法将经线性化的pZF07-16、质粒pBICP0与BHV-1基因组DNA在牛肾细胞(MDBK)上共转染,成功地进行了同源重组,7轮筛选和纯化获得了BHV-1 TK-/EGFP+突变株。
2)牛传染性鼻气管炎TK~-突变株的构建
提取BHV-1 TK-/EGFP+突变株基因组,再次以磷酸钙转染法将经线性化的pZF08-21、质粒pBICP0与基因组DNA在MDBK上共转染,同源重组去掉EGFP荧光标签,经5轮纯化成功的获得了BHV-1 TK-突变株。
3)牛传染性鼻气管炎TK-/gE-/EGFP+双缺失突变株的构建
方法同1,用PCR扩增gE基因上游和下游约1.0kb和1.1kb的片段,分别克隆于载体pcDNA3.1(+)myc-His B中。通过限制性酶切将上游同源臂回收,插入到下游的克隆载体中,构建中间转移载体,并命名为pZF09-08。然后插入EGFP报告基因,构建成重组转移质粒,并命名为pZF09-15。以磷酸钙转染法将经线性化的pZF09-15、质粒pBICP0与BHV-1 TK"缺失突变株基因组DNA在MDBK细胞上共转染,成功地进行同源重组,经过7轮筛选和纯化获得了BHV-1 TK-/gE-/EGFP+突变株。
2.牛传染性鼻气管炎TK-/gE-/EGFP+双缺失突变株的安全性和保护力研究
1)牛传染性鼻气管炎TK-/gE-/EGFP+双缺失突变株对牛的安全性研究
安全性研究包括三部分：接种试验、同居试验、再激活试验。接种试验和同居试验：将试验组按剂量分为105,106,107三栏,滴鼻免疫4X105、4X106、4X107PFU的双缺失突变株,另设空白对照组。另在各栏中同时喂养未接种的牛作为同居对照。测量试验牛的体温,观察精神状态和采食情况,采集检测样品。再激活试验：接种免疫28天后,连续3天以15mg／头的剂量肌肉注射地塞米松,以激活处于潜伏感染状态的病毒。试验证实了牛传染性鼻气管炎TK-/gE-/EGFP+双缺失突变株具有很高的安全性。
2)牛传染性鼻气管炎TK-/gE-/EGFP+双缺失突变株对牛的保护力研究
两月龄牛分别用4X105、4X106、4X107PFU的双缺失突变株免疫4周后,以4X107PFU BHV-1强毒攻击。同时设免疫DMEM的空白对照组。分别于攻毒后1d、2d、3d、5d、7d、14d、21d和28d测量体温、采集检测样品。试验结果表明：牛传染性鼻气管炎TK-/gE-/EGFP+双缺失突变株对牛具有很强的保护力。
总之,本研究表明,BHV-1 TK-/gE-可以作为候选疫苗毒株,为我国牛传染性鼻气管炎根除计划提供工具。
Infectious bovine rhinotracheitis (IBR), caused by bovine herpesvirus type 1 (BHV-1), is known as an acute, pyrexic and contagious disease. It's a major pathogen of cattle causing respiratory and genital tract infections such as bovine rhinotracheitis, conjunctivitis, fever, infectious pustular vulvovaginitis, and abortion. Since BHV-1 was discovered, IBR has occurred all over the world, and causes great economic loss to the cattle and milk industry. IBR is ranked in list B diseases by OIE, and the main measure for eradication is to slaughter the infected cattle. In addition to this, Immunization is an effective way to control it. Many live BHV-1 vaccines has been developed such as BHV-1 TK null, BHV-1 gE null and so on.
The genome of BHV-1 is a double-strand DNA of about 138 kb, with the G+C content of 72%. It contains two unique areas called the long unique (UL) and the short unique(US), and two reverse repeat units, e.t. IR and TR, flanking the US area. BHV-1 encodes 10 glycoproteins, including glycoprotein E (gE). gE is a non-essential gene for virus growth, but it influences cell-to-cell transmission in MDBK cell monolayer. On the other hand, thymidine kinase (TK) gene, another main virulence determing gene, is also non-essential for virus proliferation. It plays a very important role in the latency infection in the nervous system.
The objective of this study is to construct a bovine herpersvirus type 1 live modified vaccine strain with TK gene and gE gene deletion, providing a platform for a live viral vector expressing exogenous genes. Safety and efficacy of the both TK gene and gE gene deleted vaccine strain was carried out. The research results are summarized as follows:
1. Construction of BHV-1 TK-/gE-/EGFP+ double gene-deleted mutants
1). Construction of BHV-1 TK-/EGFP+ gene-deleted mutant
Two pairs of primers of TK gene was synthesized based on the genomic DNA sequence of BHV-1 (GenBank Accession No. AJ004801). The upstream (1.0kb) and downstream (1.1kb) of TK gene homologous arms were amplified by PCR, sequenced, and were directly cloned into pBluescriptⅡSK(+) vector one by one by restriction enzymes. Then EGFP expression cassette was inserted in between the two homologous arms. The recombinant plasmid was linearized and co-transfect into MDBK cells together with pBICPO and BHV-1 wild-type genomic DNA. By green fluorescence screening and plaque purification, the recombinant BHV-1 TK-/EGFP+ was generated.
2). Construction of BHV-1 TK- gene-deleted mutant
The pZF08-21 was linearized and co-transfected into MDBK cells together with pBICPO and BHV-1 TK-/EGFP+ genomic DNA. Through five rounds of flurescence selection and plaque purification, EGFP expression cassette was successfully removed, and BHV-1 TK- gene-deleted virus was generated.
3). Construction of BHV-1 TK-/gE-/EGFP+ double gene-deleted mutant
Two pairs of primers of gE gene was synthesized based on the genomic DNA sequence of BHV-1 (GenBank accession No. AJ004801). The upstream (1.1kb) and downstream (1.0kb) of gE gene homologous arms were amplified by PCR, sequenced, and were directly cloned into pcDNA3.1(+) myc-His B vector one by one by restriction enzymes. The EGFP expression cassette was inserted in between the two homologous arms. The transfer plasmid was linearized and co-transfected into MDBK cells together with pBICPO and BHV-1 TK- genomic DNA. Through nine rounds of flurescence selection and plaque purification, BHV-1 TK-/gE-/EGFP+ double gene-deleted mutant was generated.
2. The safety and efficacy of BHV-1 TK-/gE-/EGFP+ double gene-deleted vaccine strain in cattle
1)The safety of BHV-1 TK-/gE-/EGFP+ double gene-deleted mutant
The safety experiment was divided into three parts.They were inoculation experiment, cohabitation experiment and reactivation experiment. Three different immune dose of vaccine strain (4×105PFU,4×106PFU,4×107PFU) were infected intranasally in 10 cattle. In addition, unvaccinated cattle was designated as cohabitation test in the inoculation group. Body temperature was mesured daily, serum samples and swabs were collected. Then, dexamethasone were injected for three days to reactivate the latent virus.The safe experiment shows that the TK-/gE-/EGFP+ gene-deleted mutant is quite safe for cattle.
2)The efficacy of BHV-1 TK-/gE-/EGFP+ double gene-deleted mutant
Vaccination group and negative control group were classified in this experiment. Three different immune dose of vaccine strain (4×105PFU,4×106PFU,4×107PFU) were used to infect 9 cattle of two month old in the vaccination group, while DMEM were used in the negative control group (6 cattle). Four weeks post inoculation, both vaccination group and the control group were challenged with 4×107PFU BHV-1 wild-type virus intranasally. Body temperature was mesured daily, serum samples and swabs were collected on 1d,2d, 3d,5d,7d,14d,21d and 28d post challenging. The efficacy experiment shows that the BHV-1 TK-/gE- double gene-deleted virus could provide full immunoprotectivity to cattle.
引文
1.艾玉萍,谭诗文,冉懋韬,龙鳌,金志强,杨粤勇,王凡,滕代孝.贵州省奶牛传染性鼻气管炎的血清学调查.上海畜牧兽医通讯,2004,5：19
2.邓艳,魏冬霞,姚美兰.牛传染性鼻气管炎诊断及防治研究进展.中国动物保健,2005,6：21
3.冯斌,秦贞奎,俞太尉.中国进口牛传染性鼻气管炎检疫政策回顾.[J].中国进出境动植检,1998,4：4-5
4.葛菡,程安春,汪铭书,朱德康,罗启慧,贾仁勇,郭宇飞,陈孝跃.疱疹病毒TK基因的研究进展.中国兽医科学,2008,38：86-90
5.郭巍,朱远茂,薛飞,相文华.牛疱疹病毒1型作为或病毒载体的研究进展.中国生物工程杂志,2005,32-35
6.霍蕾,霍烽.牛传染性鼻气管炎的检疫及防控.内蒙古农业大学学报,2005,3(26)：120
7.孔繁德,徐淑菲,周斌华,陈信忠,谢明星,龚艳清,王景明,郑征,陆承平.进口奶牛牛传染性鼻气管炎病毒的分离与鉴定.中国动物检疫,2006,23(4)：29-30
8.李凯年.牛疱疹病毒1型(BHV-1)感染最新研究进展.畜牧兽医科技信息,2003,12：2-4
9.李继昌,童光志,张秀英,刘忠贵.牛传染性鼻气管炎的生物学诊断与防治.中国兽药杂志,2001,35(6)：45-48
10.李继昌,于少军,刘忠贵.牛传染性鼻气管炎病的研究进展.黑龙江畜牧兽医.2002,4：50
11.李继昌.牛传染性鼻气管炎重组疫苗的基础研究.[博士学位论文].哈尔滨：东北农业大学图书馆,2002
12.李继昌,鲁成武,殷会成.牛疱疹病毒1型活载体疫苗的构建及研究进展.东北农业大学学报,2004,35(3)：369-372
13.李兆利,薛飞,朱远茂.牛传染性鼻气管炎病毒gB蛋白研究进展.动物医学进展,2006,27(4)：1-4
14.李佐波,曹兴春.牛传染性鼻气管炎的临床诊断及预防.畜牧兽医科技信息,2006,6：46
15.刘欣宴,冯卫国,吴春涛.牛传染性鼻气管炎病的研究进展.山东科学,2006,6(19)：65-69
16.刘正飞.伪狂犬病TK-/gE-基因缺失疫苗研究.[博士学位论文].武汉：华中农业大学图书馆,2002
17.罗琼,李力施,陈茹,林志雄.从进口奶牛中分离出牛传染性鼻气管炎病毒.中国兽医杂志,2005,41(9)：45-46
18.世界动物卫生组织著.农业部畜牧兽医局译.哺乳动物、禽、蜜蜂A和B类疾病诊断试验和疫苗标准手册.北京：中国农业科学技术出版社,2002
19.孙凌志,陈庆勋,包盛,李丽.牛传染性鼻气管炎的诊断与综合疗法.国动物检疫,2005,6(22)：34
20.王海燕.IBR间接ELISA的建立及重组gE糖蛋白与单抗的研制.[博士学位论文].长春：吉林大学图书馆,2006
21.王海燕,朱远茂,薛飞,童光志,赵立平,相文华,韩文瑜.牛传染性鼻气管炎病毒gE基因的截短克隆与表达.中国预防兽医学报,2006,28(3)：289-293
22.王桂英.牛传染性鼻气管炎的流行与防制.畜牧兽医科技信息,2006,6：29
23.王延辉.牛传染性鼻气管炎病毒gC-/LacZ+基因缺失毒株的构建及gD糖蛋白的截短表达与活性检测.[硕士学位论文].哈尔滨：哈尔滨兽医研究所,2007
24.吴春涛.牛疱疹病毒Ⅰ型重组抗原的制备及间接ELISA方法的建立.[硕士学位论文].泰安：山东农业大学图书馆,2006
25.吴春涛,王建华,侯艳梅,赵祥平,霍蕾,董志珍。牛疱疹病毒I gB基因的原核表达及其抗原性分析.中国兽医科学,2006,36(07)：523-528
26.肖定汉,李兰华,江焕贤,王乐元.奶牛传染性鼻气管炎(IBR)的防治研究.中国奶牛,2004,4：43-45
27.杨春明.刚察地区牛传染性鼻气管炎的血清学调查.青海畜牧兽医杂志,2003,33(2)：39
28.颜邦芬.牛传染性鼻气管炎病毒抗体间接ELISA的建立及其在流行病学研究中的应用.[硕士学位论文].武汉：华中农业大学图书馆,2007
29.殷震,刘景华.动物病毒学(第二版).北京：科学出版社,1997
30.张桂红,童光志,王柳,仇华吉,赵晓岩,张绍杰,于力,刘宝全.牛传染性鼻气管炎病毒TK基因缺失株的构建.中国预防兽医学报,1999,21：4
31.张桂红,童光志,王柳,仇华吉,赵晓岩,张绍杰,刘宝全.牛传染性支气管炎病PCR诊断方法的研究.黑龙江畜牧兽医,2000,4：1-2
32.张伟中,张桂芬,李晓辉.牛传染性鼻气管炎的临床诊断及防制措施.畜牧兽医科技信息,2007,2：52
33.朱远茂,王海燕,薛飞,辛九庆,赵立平,童光志,郭巍,李兆利,相文华.牛传染性鼻气管炎间接ELISA诊断方法的建立.中国兽医科技,2005,35：959-963
34.庄东明.IBRVgC和gD基因的原核表达和真核表达载体的构建.[硕士学位论文].泰安：山东农业大学图书馆,2006
35. Al-Mubarak.A, J.Simon, C.Coats, J.D.Okemba, M.D.Burton, S.I.Chowdhury. Glycoprotein E (gE) specified by bovine herpesvirus tpye 5 (BHV-5) enables trans-neuronal virus spread and neurovirulence without being a structural component of enveloped virions. Virology,2007,365:398-409
36. Aykut Ozkul, Basak Demir, Taner Karaoglu, Feray alkan, Ender Dincer, Taner Oncel, and Ibrahlm Burgu. Maturation of immunoglobulin G avidity after inactive gE deleted Bovine herpesvirus type 1(BHV-1) marker vaccination. Viral Immunology, 2008,1(1):3-11
37. Beer M, Konig P, Schielke G, Trapp S. Diagnostic markers in the prevention of bovine herpesvirus type 1:possibilities and limitations. Berl. Munch. Tierarztl. Wschr,2003,116(5-6):183-191
38. Belknap EB, Walters LM., Kelling C, Ayers VK, NorrisJ, McMillen J, Hayhow C, Cochran M, Reddy DN, Wright J, Collins JK. Immunogenicity and protective effcacy of a gE, gG and US2 gene-deleted bovine herpesvirus-1 (BHV-1) vaccine. Vaccine, 1999,17(18):2297-2305
39. Benoit Muylkens, Francois Meurens, Frederic Schynts, Frederic Farnir,Aldo Pourchet, Marjorie Bardiau, Sacha Gogev, Julien Thiry, Adeline Cuisenaire, Alain Vanderplasschen and Etienne Thiry. Intraspecific bovine herpesvirus 1 recombinants carrying glycoprotein E deletion as a vaccine marker are virulent in cattle.J Virol, 2006,87:2149-2154
40. Benoit Muylkens, Francois Meurens, Frederic Schynts, Katalin de Fays, Aldo Pourchet, Julien Thiry, Alain Vanderplasschen, Nadine Antoine, Etienne Thiry. Biological characterization of bovine herpesvirus 1 recombinants possessing the vaccine glycoprotein E negative phenotype.Veterinary Microbiology,2006,113: 283-291
41. Denis M, Hanon E, Rijsewijk FA, Kaashoek MJ, van Oirschot JT, Thirty E, Pastoret PP. The role of glycoprotein gC, gE, gI and gG in the induction of cell-mediated immune responses to bovine herpesvirus 1.Vet Microbiol,1996,53(1-2):121-132
42. Galeota JA, Flores EF, Kit S, Kit M, Osorio FA. A quantitative study of the efficacy of a deletion mutant bovine herpesvirus-1 differential vaccine in reducing the establishment of latency by wildtype virus. Vaccine,1997,15(2):123-128
43. Graham, F.L. and Van der Eb, A.J. Virology,1973,52:456.
44. Hamel F, Simard C. Mapping of the Bovine Herpesvirus Glycoprotein C Promoter Region and its Spedific Transativation by the Viral BICP27 Gene Product. Archives of Virology,2003,148(1):137-152
45. Hanon E, Kei G, van Drunen Little-van den Hurk S, Griebel P, Vanderplasschen A, Rijsewijk FA, Babiuk L, Pastoret PP. Bovine herpesvirusl-induced apoptotic cell death:role of glycoprotein D. Virology,1999,257(1):191-197
46. Hutchings DL, van Drunen Little-van den Hurk S, Babiuk LA. Lymphocyte proliferative responses to separated bovine herpesvirus 1 proteins in immune cattle. J Virol,1990,64(10):5114-5122
47. Jones C. Herpes simplex virus type 1 and bovine herpesvirus 1 latency. Clin Microbiol Rev,2003,16(1):79-95
48. Jones C, V.Geiser, G.Henderson, Y.Jiang, F.Meyer,S.Perez, Y.Zhang. Functional analysis of bovine herpesvirus 1 (BHV-1) genes expressed during latency. Veterinary Microbiology,2006,113:199-210
49. Joze Grom, Peter Hostnik, Ivan toplak, Darja Barlic-Maganja. Molecular detection of BHV-1 in artificially inoculated semen and in the semen of latently infected bull treated with dexamethasone. The Veterinary Journal,2006,171:539-544
50. Kaashoek M.J, Moerman A, Madic J.A, Weerdmeester K, Maris-Veldhuis M, Rijsewijk F.A, van Oirschot J.T. An inactivated vaccine based on a Glycoprotein E-negative strain of bovine herpesvirus 1 induces protective immunity and allows serological differentiation. J. Vaccine,1995,13(4):342-346
51. Kaashoek M.J, and Van Oirschot, J.T. Early immunity induced by a live gE-negative bovine herpesvirus 1 marker vaccine. J. Vet. Microbiol,1996,53:20-26
52. Kaashoek M.J, F.A.M. Rijsewijk, R.C. Ruuls, G.M. Kcil, E. Thiry, P.P.Pastoret and J.T. Van Oirschot. Virulence,immunogenicity and reactivation of bovine herpesvirus 1 mutants with a deletion in the gC,gG,gI,gE,or in both the gI and gE gene.Vaccine, 1998,16(8):802-809
53. Kilari S, Rai A. Molecular cloning and restriction endonuclease analysis of 0.4 kb Hin dⅢ'O' fragment of bovine herpesvirus 1 DNA. Indian J Biochem Biophys,1993, 30(3):144-150
54. Kit S, Oavi H, Gaines JD, Billingsley P, McConnell S. Thymidine kinase-negative bovine herpesvirus type 1 mutant is stable and highly attenuated in calves. Arch Virol, 1985,86:63-83
55. Kit M, Kit S, Little SP, Di Marchi RD, Gale C. Bovine herpesvirus-1 (infectious bovine rhinotracheitis virus)-based viral vector which expresses foot-and-mouth disease epitopes. Vaccines,1991,9(8):564-572
56. Kit S, Otsuka H, Kit M. Expression of porcine pseudorabies virus genes by a bovine herpesvirus-1 (infectious bovine rhinotracheitis virus) vector. Arch Virol,1992,124: (1-2):1-20
57. Latchman DS, Herpes simplex virus vectors for gene delivery to a variety of different cell types. Curr Gene Ther,2002,2(4):415-426
58. Lemaire M, Schynts F, Meyer G, Georgin JP, Baranowski E, Gabriel A, Ros C, Belak S, Thiry E. Latency and reactivation of a glycoprotein E negative bovine herpesvirus type 1 vaccine:influence of virus load and effect of specific maternal antibodies. Vaccine,2001,19:4795-4804
59. Leonard J, BelloU, CharlesW, et al.Bovine herpesvirus-1 as a live virus veetor for expression of foreign genes. J. Virology,1992,190:666-673
60. Liang X.P, Babiuk L.A, Zamb T.J,et al. An in vivo study of a glyprotein gⅢ-negative bovine herpesvirus 1(BHV-1) mutant expressing β-galactosidase: evaluation of the role of gⅢ in virus infectivity and its use as a vector for mucosal immunization. J. Virology,1992,189:629-639
61. Lome A, Babiuk, Barry T.Rouse. Herpesvirus vaccines. Advanced Drug Delivery Reviews,1996,21:63-76
62. Z.F.Liu, M.C.S.Brum, A.Doster, C.Jones, and S.I.Chowdhury. A bovine herpesvirus type 1 mutant virus specifying a carboxyl-terminal truncation of glycoprotein E is defective in anterograde neuronal transport in rabbits and calves, J Virol,2008,82, 7432-7442
63. Mackett B M, Smith G L, Moss B.Vaccinia virus:a selectable eukaryotic cloning and expression vector. J. Proe. Nat 1. Acad. Sci.,1982,79:7415-7419
64. Mayfield JE, Good PJ, VanOort HJ, Campbell AR, Reed DE. Cloning and cleavage site mapping of DNA from bovine herpesvirus (strain). J Virol,1983,47(1): 259-264
65. Muralidhar S D, Thanuja C, Nagendra R, et al. Induction of cytotoxic T-lymphocytes specific for bovine herpesvirus-1 by DNA immunization. J. Vaccine,2002,20: 3744-3751
66. Muylkens B, Thiry J, Kirten P, Schynts F, Thiry E. Bovine herpesvirus infection and infectious bovine rhinotracheitis.Vet Res,2007,38(2):181-209
67. Niesalla.H.S, A.Dale, J.D.Slater, S.F.E.Scholes, J.Archer, D.J.Maskell, A.W.Tucker. Critical assessment of an in vitro bovine respiratory organ culture system:A model of bovine herpesvirus-1 infection. Journal of Virological Methods,2009,158:123-129
68. Okay Saydam, Carlos Abril, Bernd Vogt, Mathias Ackermann, MartinSchwyzer. Transactivator Protein BICP0 of Bovine Herpesvirus 1 (BHV-1) Is Blocked by Prostaglandin D2 (PGD2), Which Points to a Mechanism for PGD2-Mediated Inhibition of BHV-1 Replication. JVI,2004.78:8.3805-3810
69. Otsuka H, Xuan X. Construction of bovine herpesvirus-1 (BHV-1) recombinants which express pseudorabies virus(PRV) glycoprteins gB, gC, gD and gE. Arch Virol, 1996,141(1):57-71
70. Pagnini.U, L.De Martino, S.Montagnaro, A.Diodato, M.Longo,F.Pacelli, G.Pisanelli, G.Iovane. Bovine herpesvirus typel (BHV-1) up-regulates telomerase activity in MDBK cells. Veterinary Microbiology,2006,113:231-236
71. Patel J.R. Characteristics of live bovine herpesvirus-1 vaccines. The Veterinary Journal,2005,169:404-416
72. Rock D L, Beam S L, Mayfield J E:J. Virol.,1987,61:3827-38311
73. Sascha Trapp, Nikolaus Osterrieder, Gunther M. Keil, Martin Beer. Mutagenesis of a bovine herpesvirus type 1 genome cloned as an infectious bacterial artificial chromosome:analysis of glycoprotein E and G double deletion mutants. Virology, 2003,84:301-306
74. Scahw. Report on Bovine Herpesvirus 1 (BHV-1) marker vaccines and the accompanying diagnostic tests. A. Scientific Committee on Animal Heslth and Welfare,2000, European Commission Health & Consumer Protection Directorate-General
75. Schmitt J,Becher P, Thiel HJ, Keil GM. Expression of bovine viral diarrhoea virus glycoprotein E2 by bovine herpesvirus-1 from a synthetic ORF and incorporation of E2 into recombinant virions. J. Gen Virol,1999,80 (11):2839-2848
76. Schynts F, M.Lemaire, C.Ros, S.Belak, E.Thiry. Establishment of latency associated with glycoprotein E (gE) seroconversion after bovine herpesvirus 1 infection in calves with high levels of passive antibodies lacking gE antibodies. Veterinary Microbiology,2001,82:211-222
77. Smith GA, Young PL, Rodwell BJ, Kelly MA, Srorie GJ, Farrah CA, Mattick JS. Development and trial of a bovine herpesvirus 1-thymidine kinase deletion virus as a vaccine. Aust Vet J,1994; 71(3):65-70
78. Straub OC. Suitability of the immunodiffusion test for determining humoral antibodies to bovine herpesvirus (BHV-1). Berl Munch Tierarztl Wochenschr. German,1986,99(12):424-427
79. S van Drunen Little-van den Hurk, T Zamb, LA Babiuk. Synthesis, cellular location and immunogenicity of bovine herpesvirus-1 glycoproteins gⅠ and gⅢ expressed by recombinant vaccinia virus. J. V irol,1989,63 (5):2159-2168
80. Taylor G, Rijsewijk FA, Thomas LH, Wyld SG, Gaddum RI, Cook RS, Iorrison WI, Hensen E, van Oirschot JT, Keil G. Resistance to bovine respiratory syncytial virus (BRSV) induced in calves by a recombinant bovine herpesvirus-1 expressing the attachment glycoprotein of BRSV. J. Gen Virol,1998,79:1759-1767
81. Tikoo SK, Campos M, Babiuk LA. Bovine herpesvirus 1(BHV-1):biology, pathogenesis, and control.Adv. Virus Res,1995,45:191-223
82. Trapp S, Osterrieder N, Keil GM, Beer M. Mutagenesis of a bovine herpesvirus type 1 genome cloned as an infectious bacterial artificial chromosome:analysis of glycoprotein E and G double deletion mutants. Journal of General Virology,2003, 84:301-306
83. Todd JD,Volenec FJ, Paton IM. Intranasal vaccination against infectious bovine rhinotracheitis:Studies on early onset of protection and use of the vaccine in pregnant cows. JAVMA,1971,159(11):1370-1374
84. Thomas K R, Capecchi M R. Site-directed mutagenesis by gene targeting in mouse-embryo-derived stem cells. Cell,1987,51(3):503～512
85. Vilcek S. Detection of the bovine hepesvirus-1 (BHV-1) genome by PCR. J Vrol Methods,1993,41(2):245-257
86. Vicki Geiser, Suzanne Rose, Clinton Jones. Bovine hepesvirus type 1 induces cell death by a cell-type-dependent fashion. Microbial Pathogenesis,2008,44:459-466.
87. Yange Zhang, Joe Zhou, Clinton Jones. Identification of functional domains within the bICPO protein encoded by bovine herpesvirus 1, Journal of General Virology, 2005,86,879-886
2.邓艳,魏冬霞,姚美兰.牛传染性鼻气管炎诊断及防治研究进展.中国动物保健,2005,6：21
3.冯斌,秦贞奎,俞太尉.中国进口牛传染性鼻气管炎检疫政策回顾.[J].中国进出境动植检,1998,4：4-5
4.葛菡,程安春,汪铭书,朱德康,罗启慧,贾仁勇,郭宇飞,陈孝跃.疱疹病毒TK基因的研究进展.中国兽医科学,2008,38：86-90
5.郭巍,朱远茂,薛飞,相文华.牛疱疹病毒1型作为或病毒载体的研究进展.中国生物工程杂志,2005,32-35
6.霍蕾,霍烽.牛传染性鼻气管炎的检疫及防控.内蒙古农业大学学报,2005,3(26)：120
7.孔繁德,徐淑菲,周斌华,陈信忠,谢明星,龚艳清,王景明,郑征,陆承平.进口奶牛牛传染性鼻气管炎病毒的分离与鉴定.中国动物检疫,2006,23(4)：29-30
8.李凯年.牛疱疹病毒1型(BHV-1)感染最新研究进展.畜牧兽医科技信息,2003,12：2-4
9.李继昌,童光志,张秀英,刘忠贵.牛传染性鼻气管炎的生物学诊断与防治.中国兽药杂志,2001,35(6)：45-48
10.李继昌,于少军,刘忠贵.牛传染性鼻气管炎病的研究进展.黑龙江畜牧兽医.2002,4：50
11.李继昌.牛传染性鼻气管炎重组疫苗的基础研究.[博士学位论文].哈尔滨：东北农业大学图书馆,2002
12.李继昌,鲁成武,殷会成.牛疱疹病毒1型活载体疫苗的构建及研究进展.东北农业大学学报,2004,35(3)：369-372
13.李兆利,薛飞,朱远茂.牛传染性鼻气管炎病毒gB蛋白研究进展.动物医学进展,2006,27(4)：1-4
14.李佐波,曹兴春.牛传染性鼻气管炎的临床诊断及预防.畜牧兽医科技信息,2006,6：46
15.刘欣宴,冯卫国,吴春涛.牛传染性鼻气管炎病的研究进展.山东科学,2006,6(19)：65-69
16.刘正飞.伪狂犬病TK-/gE-基因缺失疫苗研究.[博士学位论文].武汉：华中农业大学图书馆,2002
17.罗琼,李力施,陈茹,林志雄.从进口奶牛中分离出牛传染性鼻气管炎病毒.中国兽医杂志,2005,41(9)：45-46
18.世界动物卫生组织著.农业部畜牧兽医局译.哺乳动物、禽、蜜蜂A和B类疾病诊断试验和疫苗标准手册.北京：中国农业科学技术出版社,2002
19.孙凌志,陈庆勋,包盛,李丽.牛传染性鼻气管炎的诊断与综合疗法.国动物检疫,2005,6(22)：34
20.王海燕.IBR间接ELISA的建立及重组gE糖蛋白与单抗的研制.[博士学位论文].长春：吉林大学图书馆,2006
21.王海燕,朱远茂,薛飞,童光志,赵立平,相文华,韩文瑜.牛传染性鼻气管炎病毒gE基因的截短克隆与表达.中国预防兽医学报,2006,28(3)：289-293
22.王桂英.牛传染性鼻气管炎的流行与防制.畜牧兽医科技信息,2006,6：29
23.王延辉.牛传染性鼻气管炎病毒gC-/LacZ+基因缺失毒株的构建及gD糖蛋白的截短表达与活性检测.[硕士学位论文].哈尔滨：哈尔滨兽医研究所,2007
24.吴春涛.牛疱疹病毒Ⅰ型重组抗原的制备及间接ELISA方法的建立.[硕士学位论文].泰安：山东农业大学图书馆,2006
25.吴春涛,王建华,侯艳梅,赵祥平,霍蕾,董志珍。牛疱疹病毒I gB基因的原核表达及其抗原性分析.中国兽医科学,2006,36(07)：523-528
26.肖定汉,李兰华,江焕贤,王乐元.奶牛传染性鼻气管炎(IBR)的防治研究.中国奶牛,2004,4：43-45
27.杨春明.刚察地区牛传染性鼻气管炎的血清学调查.青海畜牧兽医杂志,2003,33(2)：39
28.颜邦芬.牛传染性鼻气管炎病毒抗体间接ELISA的建立及其在流行病学研究中的应用.[硕士学位论文].武汉：华中农业大学图书馆,2007
29.殷震,刘景华.动物病毒学(第二版).北京：科学出版社,1997
30.张桂红,童光志,王柳,仇华吉,赵晓岩,张绍杰,于力,刘宝全.牛传染性鼻气管炎病毒TK基因缺失株的构建.中国预防兽医学报,1999,21：4
31.张桂红,童光志,王柳,仇华吉,赵晓岩,张绍杰,刘宝全.牛传染性支气管炎病PCR诊断方法的研究.黑龙江畜牧兽医,2000,4：1-2
32.张伟中,张桂芬,李晓辉.牛传染性鼻气管炎的临床诊断及防制措施.畜牧兽医科技信息,2007,2：52
33.朱远茂,王海燕,薛飞,辛九庆,赵立平,童光志,郭巍,李兆利,相文华.牛传染性鼻气管炎间接ELISA诊断方法的建立.中国兽医科技,2005,35：959-963
34.庄东明.IBRVgC和gD基因的原核表达和真核表达载体的构建.[硕士学位论文].泰安：山东农业大学图书馆,2006
35. Al-Mubarak.A, J.Simon, C.Coats, J.D.Okemba, M.D.Burton, S.I.Chowdhury. Glycoprotein E (gE) specified by bovine herpesvirus tpye 5 (BHV-5) enables trans-neuronal virus spread and neurovirulence without being a structural component of enveloped virions. Virology,2007,365:398-409
36. Aykut Ozkul, Basak Demir, Taner Karaoglu, Feray alkan, Ender Dincer, Taner Oncel, and Ibrahlm Burgu. Maturation of immunoglobulin G avidity after inactive gE deleted Bovine herpesvirus type 1(BHV-1) marker vaccination. Viral Immunology, 2008,1(1):3-11
37. Beer M, Konig P, Schielke G, Trapp S. Diagnostic markers in the prevention of bovine herpesvirus type 1:possibilities and limitations. Berl. Munch. Tierarztl. Wschr,2003,116(5-6):183-191
38. Belknap EB, Walters LM., Kelling C, Ayers VK, NorrisJ, McMillen J, Hayhow C, Cochran M, Reddy DN, Wright J, Collins JK. Immunogenicity and protective effcacy of a gE, gG and US2 gene-deleted bovine herpesvirus-1 (BHV-1) vaccine. Vaccine, 1999,17(18):2297-2305
39. Benoit Muylkens, Francois Meurens, Frederic Schynts, Frederic Farnir,Aldo Pourchet, Marjorie Bardiau, Sacha Gogev, Julien Thiry, Adeline Cuisenaire, Alain Vanderplasschen and Etienne Thiry. Intraspecific bovine herpesvirus 1 recombinants carrying glycoprotein E deletion as a vaccine marker are virulent in cattle.J Virol, 2006,87:2149-2154
40. Benoit Muylkens, Francois Meurens, Frederic Schynts, Katalin de Fays, Aldo Pourchet, Julien Thiry, Alain Vanderplasschen, Nadine Antoine, Etienne Thiry. Biological characterization of bovine herpesvirus 1 recombinants possessing the vaccine glycoprotein E negative phenotype.Veterinary Microbiology,2006,113: 283-291
41. Denis M, Hanon E, Rijsewijk FA, Kaashoek MJ, van Oirschot JT, Thirty E, Pastoret PP. The role of glycoprotein gC, gE, gI and gG in the induction of cell-mediated immune responses to bovine herpesvirus 1.Vet Microbiol,1996,53(1-2):121-132
42. Galeota JA, Flores EF, Kit S, Kit M, Osorio FA. A quantitative study of the efficacy of a deletion mutant bovine herpesvirus-1 differential vaccine in reducing the establishment of latency by wildtype virus. Vaccine,1997,15(2):123-128
43. Graham, F.L. and Van der Eb, A.J. Virology,1973,52:456.
44. Hamel F, Simard C. Mapping of the Bovine Herpesvirus Glycoprotein C Promoter Region and its Spedific Transativation by the Viral BICP27 Gene Product. Archives of Virology,2003,148(1):137-152
45. Hanon E, Kei G, van Drunen Little-van den Hurk S, Griebel P, Vanderplasschen A, Rijsewijk FA, Babiuk L, Pastoret PP. Bovine herpesvirusl-induced apoptotic cell death:role of glycoprotein D. Virology,1999,257(1):191-197
46. Hutchings DL, van Drunen Little-van den Hurk S, Babiuk LA. Lymphocyte proliferative responses to separated bovine herpesvirus 1 proteins in immune cattle. J Virol,1990,64(10):5114-5122
47. Jones C. Herpes simplex virus type 1 and bovine herpesvirus 1 latency. Clin Microbiol Rev,2003,16(1):79-95
48. Jones C, V.Geiser, G.Henderson, Y.Jiang, F.Meyer,S.Perez, Y.Zhang. Functional analysis of bovine herpesvirus 1 (BHV-1) genes expressed during latency. Veterinary Microbiology,2006,113:199-210
49. Joze Grom, Peter Hostnik, Ivan toplak, Darja Barlic-Maganja. Molecular detection of BHV-1 in artificially inoculated semen and in the semen of latently infected bull treated with dexamethasone. The Veterinary Journal,2006,171:539-544
50. Kaashoek M.J, Moerman A, Madic J.A, Weerdmeester K, Maris-Veldhuis M, Rijsewijk F.A, van Oirschot J.T. An inactivated vaccine based on a Glycoprotein E-negative strain of bovine herpesvirus 1 induces protective immunity and allows serological differentiation. J. Vaccine,1995,13(4):342-346
51. Kaashoek M.J, and Van Oirschot, J.T. Early immunity induced by a live gE-negative bovine herpesvirus 1 marker vaccine. J. Vet. Microbiol,1996,53:20-26
52. Kaashoek M.J, F.A.M. Rijsewijk, R.C. Ruuls, G.M. Kcil, E. Thiry, P.P.Pastoret and J.T. Van Oirschot. Virulence,immunogenicity and reactivation of bovine herpesvirus 1 mutants with a deletion in the gC,gG,gI,gE,or in both the gI and gE gene.Vaccine, 1998,16(8):802-809
53. Kilari S, Rai A. Molecular cloning and restriction endonuclease analysis of 0.4 kb Hin dⅢ'O' fragment of bovine herpesvirus 1 DNA. Indian J Biochem Biophys,1993, 30(3):144-150
54. Kit S, Oavi H, Gaines JD, Billingsley P, McConnell S. Thymidine kinase-negative bovine herpesvirus type 1 mutant is stable and highly attenuated in calves. Arch Virol, 1985,86:63-83
55. Kit M, Kit S, Little SP, Di Marchi RD, Gale C. Bovine herpesvirus-1 (infectious bovine rhinotracheitis virus)-based viral vector which expresses foot-and-mouth disease epitopes. Vaccines,1991,9(8):564-572
56. Kit S, Otsuka H, Kit M. Expression of porcine pseudorabies virus genes by a bovine herpesvirus-1 (infectious bovine rhinotracheitis virus) vector. Arch Virol,1992,124: (1-2):1-20
57. Latchman DS, Herpes simplex virus vectors for gene delivery to a variety of different cell types. Curr Gene Ther,2002,2(4):415-426
58. Lemaire M, Schynts F, Meyer G, Georgin JP, Baranowski E, Gabriel A, Ros C, Belak S, Thiry E. Latency and reactivation of a glycoprotein E negative bovine herpesvirus type 1 vaccine:influence of virus load and effect of specific maternal antibodies. Vaccine,2001,19:4795-4804
59. Leonard J, BelloU, CharlesW, et al.Bovine herpesvirus-1 as a live virus veetor for expression of foreign genes. J. Virology,1992,190:666-673
60. Liang X.P, Babiuk L.A, Zamb T.J,et al. An in vivo study of a glyprotein gⅢ-negative bovine herpesvirus 1(BHV-1) mutant expressing β-galactosidase: evaluation of the role of gⅢ in virus infectivity and its use as a vector for mucosal immunization. J. Virology,1992,189:629-639
61. Lome A, Babiuk, Barry T.Rouse. Herpesvirus vaccines. Advanced Drug Delivery Reviews,1996,21:63-76
62. Z.F.Liu, M.C.S.Brum, A.Doster, C.Jones, and S.I.Chowdhury. A bovine herpesvirus type 1 mutant virus specifying a carboxyl-terminal truncation of glycoprotein E is defective in anterograde neuronal transport in rabbits and calves, J Virol,2008,82, 7432-7442
63. Mackett B M, Smith G L, Moss B.Vaccinia virus:a selectable eukaryotic cloning and expression vector. J. Proe. Nat 1. Acad. Sci.,1982,79:7415-7419
64. Mayfield JE, Good PJ, VanOort HJ, Campbell AR, Reed DE. Cloning and cleavage site mapping of DNA from bovine herpesvirus (strain). J Virol,1983,47(1): 259-264
65. Muralidhar S D, Thanuja C, Nagendra R, et al. Induction of cytotoxic T-lymphocytes specific for bovine herpesvirus-1 by DNA immunization. J. Vaccine,2002,20: 3744-3751
66. Muylkens B, Thiry J, Kirten P, Schynts F, Thiry E. Bovine herpesvirus infection and infectious bovine rhinotracheitis.Vet Res,2007,38(2):181-209
67. Niesalla.H.S, A.Dale, J.D.Slater, S.F.E.Scholes, J.Archer, D.J.Maskell, A.W.Tucker. Critical assessment of an in vitro bovine respiratory organ culture system:A model of bovine herpesvirus-1 infection. Journal of Virological Methods,2009,158:123-129
68. Okay Saydam, Carlos Abril, Bernd Vogt, Mathias Ackermann, MartinSchwyzer. Transactivator Protein BICP0 of Bovine Herpesvirus 1 (BHV-1) Is Blocked by Prostaglandin D2 (PGD2), Which Points to a Mechanism for PGD2-Mediated Inhibition of BHV-1 Replication. JVI,2004.78:8.3805-3810
69. Otsuka H, Xuan X. Construction of bovine herpesvirus-1 (BHV-1) recombinants which express pseudorabies virus(PRV) glycoprteins gB, gC, gD and gE. Arch Virol, 1996,141(1):57-71
70. Pagnini.U, L.De Martino, S.Montagnaro, A.Diodato, M.Longo,F.Pacelli, G.Pisanelli, G.Iovane. Bovine herpesvirus typel (BHV-1) up-regulates telomerase activity in MDBK cells. Veterinary Microbiology,2006,113:231-236
71. Patel J.R. Characteristics of live bovine herpesvirus-1 vaccines. The Veterinary Journal,2005,169:404-416
72. Rock D L, Beam S L, Mayfield J E:J. Virol.,1987,61:3827-38311
73. Sascha Trapp, Nikolaus Osterrieder, Gunther M. Keil, Martin Beer. Mutagenesis of a bovine herpesvirus type 1 genome cloned as an infectious bacterial artificial chromosome:analysis of glycoprotein E and G double deletion mutants. Virology, 2003,84:301-306
74. Scahw. Report on Bovine Herpesvirus 1 (BHV-1) marker vaccines and the accompanying diagnostic tests. A. Scientific Committee on Animal Heslth and Welfare,2000, European Commission Health & Consumer Protection Directorate-General
75. Schmitt J,Becher P, Thiel HJ, Keil GM. Expression of bovine viral diarrhoea virus glycoprotein E2 by bovine herpesvirus-1 from a synthetic ORF and incorporation of E2 into recombinant virions. J. Gen Virol,1999,80 (11):2839-2848
76. Schynts F, M.Lemaire, C.Ros, S.Belak, E.Thiry. Establishment of latency associated with glycoprotein E (gE) seroconversion after bovine herpesvirus 1 infection in calves with high levels of passive antibodies lacking gE antibodies. Veterinary Microbiology,2001,82:211-222
77. Smith GA, Young PL, Rodwell BJ, Kelly MA, Srorie GJ, Farrah CA, Mattick JS. Development and trial of a bovine herpesvirus 1-thymidine kinase deletion virus as a vaccine. Aust Vet J,1994; 71(3):65-70
78. Straub OC. Suitability of the immunodiffusion test for determining humoral antibodies to bovine herpesvirus (BHV-1). Berl Munch Tierarztl Wochenschr. German,1986,99(12):424-427
79. S van Drunen Little-van den Hurk, T Zamb, LA Babiuk. Synthesis, cellular location and immunogenicity of bovine herpesvirus-1 glycoproteins gⅠ and gⅢ expressed by recombinant vaccinia virus. J. V irol,1989,63 (5):2159-2168
80. Taylor G, Rijsewijk FA, Thomas LH, Wyld SG, Gaddum RI, Cook RS, Iorrison WI, Hensen E, van Oirschot JT, Keil G. Resistance to bovine respiratory syncytial virus (BRSV) induced in calves by a recombinant bovine herpesvirus-1 expressing the attachment glycoprotein of BRSV. J. Gen Virol,1998,79:1759-1767
81. Tikoo SK, Campos M, Babiuk LA. Bovine herpesvirus 1(BHV-1):biology, pathogenesis, and control.Adv. Virus Res,1995,45:191-223
82. Trapp S, Osterrieder N, Keil GM, Beer M. Mutagenesis of a bovine herpesvirus type 1 genome cloned as an infectious bacterial artificial chromosome:analysis of glycoprotein E and G double deletion mutants. Journal of General Virology,2003, 84:301-306
83. Todd JD,Volenec FJ, Paton IM. Intranasal vaccination against infectious bovine rhinotracheitis:Studies on early onset of protection and use of the vaccine in pregnant cows. JAVMA,1971,159(11):1370-1374
84. Thomas K R, Capecchi M R. Site-directed mutagenesis by gene targeting in mouse-embryo-derived stem cells. Cell,1987,51(3):503～512
85. Vilcek S. Detection of the bovine hepesvirus-1 (BHV-1) genome by PCR. J Vrol Methods,1993,41(2):245-257
86. Vicki Geiser, Suzanne Rose, Clinton Jones. Bovine hepesvirus type 1 induces cell death by a cell-type-dependent fashion. Microbial Pathogenesis,2008,44:459-466.
87. Yange Zhang, Joe Zhou, Clinton Jones. Identification of functional domains within the bICPO protein encoded by bovine herpesvirus 1, Journal of General Virology, 2005,86,879-886