Renin和IL-10基因与猪妊娠期的相关性研究
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摘要
妊娠期是猪重要的繁殖性状之一。在生产过程中缩短母猪的妊娠期可以缩短其繁殖周期,提高母猪的繁殖效率,降低生产成本。妊娠期是数量性状,由多基因的控制的,且单个基因的效应值较小,易受环境因素的影响,用常规的育种方法对其进行遗传改良进展缓慢。
有研究报道影响猪妊娠期的QTL位于猪的第1、9和15号染色体上,根据猪-人基因组比较图谱和基因的生理生化功能选取了两个候选基因:肾素(Renin)和白介素-10(IL-10)。应用生物信息学和比较基因组的方法对这两个基因进行了克隆,经不同品种间序列的比对发现这两个基因的突变位点,并选择三个突变位点与妊娠期和产仔数进行相关性分析:
1.根据与人的Renin基因有较高同源性的猪EST以及人和小鼠等物种该基因cDNA相似性较高的区域设计了5对引物,以梅山猪cDNA作为模板扩增、克隆并测序,拼接后获得1333 bp的整合cDNA序列,包含全部CDS和部分5'和3'非翻译区。利用获得的cDNA序列设计18对引物对其DNA序列进行扩增,克隆、测序和拼接后得到了13332bp的DNA序列。
2.利用Clustal X、ORF Finder、Spidey、MEGA4.0、PSORTⅡ等软件对猪Renin基因结构、编码的蛋白质分子量、等电点、系统进化树等特征进行了预测和分析。我们发现该基因共有9个外显子和8个内含子且内含子的剪切方式符合“GT-AG”法则,进化树分析得出该基因与反刍动物的相似性最高,与啮齿类的次之,与灵长类的最低。
3.根据猪IL-10基因的cDNA序列设计7对引物,克隆得到了该基因4062 bp的DNA序列,经过对该基因的结构进行分析,该基因包含有4个内含子,内含子的剪切方式符合“GT-AG”法则。
4.根据已获得的Renin基因的cDNA序列设计引物,用RT-PCR的方法检测其在不同组织中的表达情况,发现在肾、卵巢和输卵管中有表达,在肾中的表达最高。
5.根据获得的这两个基因的序列,清平猪、大白猪、长白猪和杜洛克猪的DNA作为模板进行扩增、克隆并测序。经序列比对,发现了在Renin的第6内含子228bp处发现一个C/T转换,第7内含子的114bp处发现一个A/G转换和IL-10基因第3内含子的835bp处发现一个A/G转换,据此建立适宜的PCR-RFLP SNP分型方法。
6.Renin的第6内含子内的突变可以被HhaⅠ酶切,建立该基因的PCR-HhaⅠ-RFLP检测方法,对该位点在清平猪、新清平猪、长白猪、大白猪,杜洛克猪和中国瘦肉猪新品系DⅣ_1系、DⅣ_2系7个中外猪群体中进行基因型分型,分析了它们的基因型频率和基因频率,并与妊娠期和产仔数进行相关性分析,结果表明该位点TT基因型在国外猪种中占有优势,地方品种和培育品种与外种猪的基因型频率之间均存在显著地差异,且都达到了极显著的水平。该位点在新清平猪中对于妊娠期的显性效应达到了显著地水平。在DⅣ_1系中CC基因型的妊娠期要显著高于TT和CT基因型的妊娠期。该位点在不同的群体中对于产仔数的加性效应和显性效应均不显著。
7.Renin基因第7内含子内的突变可以被bstxⅠ酶切,建立该基因的PCR-bstxⅠ-RFLP检测方法,在清平猪、新清平猪、长白猪、大白猪杜洛克猪和中国瘦肉猪新品系DⅣ_1系、DⅣ_2系7个中外猪群体中进行基因型分型,分析了它们的基因型频率和基因频率,并与妊娠期和产仔数进行相关性分析,结果表明该位点A等位基因在各个猪群中占有优势,在杜洛克猪中除外。大部分猪群之间基因型频率差异显著。在新清平猪中该位点对于妊娠期的显性效应差异显著,为0.93d。该位点在杜洛克猪中对于产仔数的加性效应达到极显著地水平,GG基因型的产仔数要显著地高于AG和AA基因型的产仔数。
8.IL-10的第3内含子内的突变可以被Eco81Ⅰ酶切,建立该基因的PCR-Eco81Ⅰ-RFLP检测方法,在清平猪、新清平猪、长白猪、大白猪杜洛克猪和中国瘦肉猪新品系DⅣ_1系、DⅣ_2系7个中外猪群体中进行基因型分型,分析了它们的基因型频率和基因频率,并与妊娠期进行相关性分析,结果表明该位点GG基因型在清平猪、新清平猪和杜洛克猪中占有优势,而在长白猪、大白猪、DⅣ_1系和DⅣ_2系中AA基因型占有优势。大部分猪群之间基因型频率差异显著。该位点在杜洛克猪中对于妊娠期的显性效应显著,致使AG基因型的妊娠期要显著长于AA基因型的妊娠期。该位点在大白猪中对于产仔数的加性效应显著,GG基因型的产仔数要显著少于AG和AA基因型的产仔数。
The gestation length serves as one of the most importent reproductive traits. Reducing the gestation length,taking swine breeding into account,directly results in a shorter reproductive cycle;correspondingly reduce the production cost by increasing the reproductive efficiency of sows.Gestation length is a quantitative trait which is affected by several genes' cooperation.Due to the amount of genes and every single effect it provides,as well as environmental factors,regular breeding methods have scarcely any apparent genetic improvement on this character.
The QTL related with gestation length is located at the 1,9,15 chromosome in sus scrofa.According to swine- human genome comparison and both physiological and biochemical function,two candidate genes renin and interleukin-10 have been selected in this experiment.In this experiment several methods like bioinformatics and conservation of linkage with swine and man are being applied to clone these genes and polymorphic sites have been made in order to analyze the relativity with the gestation length and and total number of piglet born.
1.Based on swine's EST which has a high homology with human renin gene and the cDNA of other species like mice,five primers have been designed on the sequences with great similarity within several animals.Meishan's cDNA has been chosen as the template which is sequenced and cloned.After splicing and integrating,the cDNA, 1333 base pair,has been acquired including the whole CDS and parts of 5' and 3' untranslationed region.Later,the DNA sequence has amplified,cloned and sequenced which was examined as 13332 base pairs.
2.With the help of software Clustal X、ORF Finder、Spidey、MEGA4.0、PSORTⅡ, renin's gene structure,the structure of protein,property,the phylogenetics tree,has been predicted and analyzed.We find that this gene with 9 extrons and 8 introns matching the GT-AG rule,is extremely resemble with rumen animals,less similar with rodents,and least conformity with primate.
3.Relied on cDNA of IL-10 gene,seven primers have designed and the whole gene has been cloned with 4062bp.After analyzing the structure of the gene,4 introns have been included and complied with the GT-AG rule.
4.RT-PCR has been used to exam renin expression in different tissues,the results demonstrate that kidney,ovaries,oviduct have expressed this gene with the highest one founded in kidney.
5.Different DNA of Qingping,Large white,Landrace and Duroc have been amplified, cloned and sequenced.Transvertion of C/T at the 6 intron,288bp,and A/G at the 7 intron 114bp in renin as well as the transvertion A/G in the third intron of IL-10 at 835bp have been found based on sequences comparison.Appropriate PCR-RFLP SNP typing methods have been established.
6.The mutation in the sixth intron of renin can be cut by enzyme HhaⅠ.Qingping,New Qingping,large white,landerace,Duroc and Chinese lean meat pig strain DⅣ_1、DⅣ_2,altogether seven kinds of swine have been typed.The genetype frequency and gene frequency were analyzed in order to dig out the relativity with gestation length and and total number of piglet born.The result shows that the specific site with genetype TT plays more prominent effect in foreign pigs,whereas local breed and cultivated strain has significant variation from foreign swine even achieved the extremely significant variation.The dominant effect on gestation length in this site in Qingping has reached significant level as well.In the DⅣ_1 strain,hogs' gestation length with gene type CC were significantly longer than that of TT and CT gene type. The dominant effect and the additive effect on total number of piglet born in this site in all the groups are not significant.
7.The mutation in seventh intron of renin can be cut by enzyme bstxⅠ.Qingping,New Qingping,large white,landerace,Duroc and Chinese lean meat pig strain DⅣ_1、DⅣ_2,altogether seven kinds of swine have been typed.The genetype frequency and gene frequency were analyzed in order to dig out the relativity with gestation length and total number of piglet born.The result shows that allele gene A has a more conspicuous advantage in any swinery,besides duroc.The gene frequency has significant variation and the dominant effect on gestation length in Qingping was significant,0.93day.The additive effect on total number of piglet born in this site in Duroc has reached significant level,hogs' total number of piglet born with gene type GG were significantly more than that of AG and AA gene type.
8.The mutation in third intron of IL-10 can be cut by enzyme Eco81Ⅰ.Qingping,New Qingping,large white,landerace,Duroc and Chinese lean meat pig strain DⅣ_1、DⅣ_2,altogether seven kinds of swine have been typed.The genetype frequency and gene frequency were analyzed in order to dig out the relativity with gestation length and total number of piglet born.The result shows that the specific site with gene type GG plays more prominent effect in Qingping,New Qingping and duroc,whereas within large white,landerace,DⅣ_1,and DⅣ_2 AA acted a more important role.Most swinery has significant variation from each other.The dominant effect on gestation length in this site in duroc has reached significant level,consequently pigs with gene type AG have a longer gestation length than those of AA gene type.The additive effect on total number of piglet born in this site in large white has reached significant level,hogs' total number of piglet bom with gene type GG were significantly fewer than that of AG and AA gene type.
有研究报道影响猪妊娠期的QTL位于猪的第1、9和15号染色体上,根据猪-人基因组比较图谱和基因的生理生化功能选取了两个候选基因:肾素(Renin)和白介素-10(IL-10)。应用生物信息学和比较基因组的方法对这两个基因进行了克隆,经不同品种间序列的比对发现这两个基因的突变位点,并选择三个突变位点与妊娠期和产仔数进行相关性分析:
1.根据与人的Renin基因有较高同源性的猪EST以及人和小鼠等物种该基因cDNA相似性较高的区域设计了5对引物,以梅山猪cDNA作为模板扩增、克隆并测序,拼接后获得1333 bp的整合cDNA序列,包含全部CDS和部分5'和3'非翻译区。利用获得的cDNA序列设计18对引物对其DNA序列进行扩增,克隆、测序和拼接后得到了13332bp的DNA序列。
2.利用Clustal X、ORF Finder、Spidey、MEGA4.0、PSORTⅡ等软件对猪Renin基因结构、编码的蛋白质分子量、等电点、系统进化树等特征进行了预测和分析。我们发现该基因共有9个外显子和8个内含子且内含子的剪切方式符合“GT-AG”法则,进化树分析得出该基因与反刍动物的相似性最高,与啮齿类的次之,与灵长类的最低。
3.根据猪IL-10基因的cDNA序列设计7对引物,克隆得到了该基因4062 bp的DNA序列,经过对该基因的结构进行分析,该基因包含有4个内含子,内含子的剪切方式符合“GT-AG”法则。
4.根据已获得的Renin基因的cDNA序列设计引物,用RT-PCR的方法检测其在不同组织中的表达情况,发现在肾、卵巢和输卵管中有表达,在肾中的表达最高。
5.根据获得的这两个基因的序列,清平猪、大白猪、长白猪和杜洛克猪的DNA作为模板进行扩增、克隆并测序。经序列比对,发现了在Renin的第6内含子228bp处发现一个C/T转换,第7内含子的114bp处发现一个A/G转换和IL-10基因第3内含子的835bp处发现一个A/G转换,据此建立适宜的PCR-RFLP SNP分型方法。
6.Renin的第6内含子内的突变可以被HhaⅠ酶切,建立该基因的PCR-HhaⅠ-RFLP检测方法,对该位点在清平猪、新清平猪、长白猪、大白猪,杜洛克猪和中国瘦肉猪新品系DⅣ_1系、DⅣ_2系7个中外猪群体中进行基因型分型,分析了它们的基因型频率和基因频率,并与妊娠期和产仔数进行相关性分析,结果表明该位点TT基因型在国外猪种中占有优势,地方品种和培育品种与外种猪的基因型频率之间均存在显著地差异,且都达到了极显著的水平。该位点在新清平猪中对于妊娠期的显性效应达到了显著地水平。在DⅣ_1系中CC基因型的妊娠期要显著高于TT和CT基因型的妊娠期。该位点在不同的群体中对于产仔数的加性效应和显性效应均不显著。
7.Renin基因第7内含子内的突变可以被bstxⅠ酶切,建立该基因的PCR-bstxⅠ-RFLP检测方法,在清平猪、新清平猪、长白猪、大白猪杜洛克猪和中国瘦肉猪新品系DⅣ_1系、DⅣ_2系7个中外猪群体中进行基因型分型,分析了它们的基因型频率和基因频率,并与妊娠期和产仔数进行相关性分析,结果表明该位点A等位基因在各个猪群中占有优势,在杜洛克猪中除外。大部分猪群之间基因型频率差异显著。在新清平猪中该位点对于妊娠期的显性效应差异显著,为0.93d。该位点在杜洛克猪中对于产仔数的加性效应达到极显著地水平,GG基因型的产仔数要显著地高于AG和AA基因型的产仔数。
8.IL-10的第3内含子内的突变可以被Eco81Ⅰ酶切,建立该基因的PCR-Eco81Ⅰ-RFLP检测方法,在清平猪、新清平猪、长白猪、大白猪杜洛克猪和中国瘦肉猪新品系DⅣ_1系、DⅣ_2系7个中外猪群体中进行基因型分型,分析了它们的基因型频率和基因频率,并与妊娠期进行相关性分析,结果表明该位点GG基因型在清平猪、新清平猪和杜洛克猪中占有优势,而在长白猪、大白猪、DⅣ_1系和DⅣ_2系中AA基因型占有优势。大部分猪群之间基因型频率差异显著。该位点在杜洛克猪中对于妊娠期的显性效应显著,致使AG基因型的妊娠期要显著长于AA基因型的妊娠期。该位点在大白猪中对于产仔数的加性效应显著,GG基因型的产仔数要显著少于AG和AA基因型的产仔数。
The gestation length serves as one of the most importent reproductive traits. Reducing the gestation length,taking swine breeding into account,directly results in a shorter reproductive cycle;correspondingly reduce the production cost by increasing the reproductive efficiency of sows.Gestation length is a quantitative trait which is affected by several genes' cooperation.Due to the amount of genes and every single effect it provides,as well as environmental factors,regular breeding methods have scarcely any apparent genetic improvement on this character.
The QTL related with gestation length is located at the 1,9,15 chromosome in sus scrofa.According to swine- human genome comparison and both physiological and biochemical function,two candidate genes renin and interleukin-10 have been selected in this experiment.In this experiment several methods like bioinformatics and conservation of linkage with swine and man are being applied to clone these genes and polymorphic sites have been made in order to analyze the relativity with the gestation length and and total number of piglet born.
1.Based on swine's EST which has a high homology with human renin gene and the cDNA of other species like mice,five primers have been designed on the sequences with great similarity within several animals.Meishan's cDNA has been chosen as the template which is sequenced and cloned.After splicing and integrating,the cDNA, 1333 base pair,has been acquired including the whole CDS and parts of 5' and 3' untranslationed region.Later,the DNA sequence has amplified,cloned and sequenced which was examined as 13332 base pairs.
2.With the help of software Clustal X、ORF Finder、Spidey、MEGA4.0、PSORTⅡ, renin's gene structure,the structure of protein,property,the phylogenetics tree,has been predicted and analyzed.We find that this gene with 9 extrons and 8 introns matching the GT-AG rule,is extremely resemble with rumen animals,less similar with rodents,and least conformity with primate.
3.Relied on cDNA of IL-10 gene,seven primers have designed and the whole gene has been cloned with 4062bp.After analyzing the structure of the gene,4 introns have been included and complied with the GT-AG rule.
4.RT-PCR has been used to exam renin expression in different tissues,the results demonstrate that kidney,ovaries,oviduct have expressed this gene with the highest one founded in kidney.
5.Different DNA of Qingping,Large white,Landrace and Duroc have been amplified, cloned and sequenced.Transvertion of C/T at the 6 intron,288bp,and A/G at the 7 intron 114bp in renin as well as the transvertion A/G in the third intron of IL-10 at 835bp have been found based on sequences comparison.Appropriate PCR-RFLP SNP typing methods have been established.
6.The mutation in the sixth intron of renin can be cut by enzyme HhaⅠ.Qingping,New Qingping,large white,landerace,Duroc and Chinese lean meat pig strain DⅣ_1、DⅣ_2,altogether seven kinds of swine have been typed.The genetype frequency and gene frequency were analyzed in order to dig out the relativity with gestation length and and total number of piglet born.The result shows that the specific site with genetype TT plays more prominent effect in foreign pigs,whereas local breed and cultivated strain has significant variation from foreign swine even achieved the extremely significant variation.The dominant effect on gestation length in this site in Qingping has reached significant level as well.In the DⅣ_1 strain,hogs' gestation length with gene type CC were significantly longer than that of TT and CT gene type. The dominant effect and the additive effect on total number of piglet born in this site in all the groups are not significant.
7.The mutation in seventh intron of renin can be cut by enzyme bstxⅠ.Qingping,New Qingping,large white,landerace,Duroc and Chinese lean meat pig strain DⅣ_1、DⅣ_2,altogether seven kinds of swine have been typed.The genetype frequency and gene frequency were analyzed in order to dig out the relativity with gestation length and total number of piglet born.The result shows that allele gene A has a more conspicuous advantage in any swinery,besides duroc.The gene frequency has significant variation and the dominant effect on gestation length in Qingping was significant,0.93day.The additive effect on total number of piglet born in this site in Duroc has reached significant level,hogs' total number of piglet born with gene type GG were significantly more than that of AG and AA gene type.
8.The mutation in third intron of IL-10 can be cut by enzyme Eco81Ⅰ.Qingping,New Qingping,large white,landerace,Duroc and Chinese lean meat pig strain DⅣ_1、DⅣ_2,altogether seven kinds of swine have been typed.The genetype frequency and gene frequency were analyzed in order to dig out the relativity with gestation length and total number of piglet born.The result shows that the specific site with gene type GG plays more prominent effect in Qingping,New Qingping and duroc,whereas within large white,landerace,DⅣ_1,and DⅣ_2 AA acted a more important role.Most swinery has significant variation from each other.The dominant effect on gestation length in this site in duroc has reached significant level,consequently pigs with gene type AG have a longer gestation length than those of AA gene type.The additive effect on total number of piglet born in this site in large white has reached significant level,hogs' total number of piglet bom with gene type GG were significantly fewer than that of AG and AA gene type.
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20. Brown AG, Leite RS, Strauss JF 3rd. Mechanisms underlying "functional"progesterone withdrawal at parturition. Ann N Y Acad Sci. 2004; 1034:36-1049
21. Casey ML, Cox SM, Word RA, MacDonald PC. Cytokines and infection-induced preterm labour. Reprod Fertil Dev. 1990; 2(5): 499-509
22. Chase AJ, Bond M, Crook MF, Newby AC.Role of nuclear factor-kappa B activation in metalloproteinase-1, -3, and -9 secretion by human macrophages in vitro and rabbit foam cells produced in vivo. Arterioscler Thromb Vasc Biol. 2002;22(5): 765-771
23. Christiaens I, Zaragoza DB, Guilbert L, Robertson SA, Mitchell BF, Olson DM.Inflammatory processes in preterm and term parturition. J Reprod Immunol. 2008;79(1): 50-57
24. Christiaens I, Zaragoza DB, Guilbert L, Robertson SA, Mitchell BF, Olson DM.Inflammatory processes in preterm and term parturition. J Reprod Immunol. 2008;79(1): 50-57
25. Condon JC, Hardy DB, Kovaric K, Mendelson CR.Up-regulation of the progesterone receptor (PR)-C isoform in laboring myometrium by activation of nuclear factor-kappaB may contribute to the onset of labor through inhibition of PR function. Mol Endocrinol. 2006; 20(4): 764-775
26. Condon JC, Jeyasuria P, Faust JM, Mendelson CR. Surfactant protein secreted by the maturing mouse fetal lung acts as a hormone that signals the initiation of parturition. Proc Natl Acad Sci U S A. 2004 ; 101(14): 4978-83
27. Cooper AC, Robinson G, Vinson GP, Cheung WT, Broughton Pipkin F. The localization and expression of the renin-angiotensin system in the human placenta throughout pregnancy.Placenta. 1999; 20(5-6): 467-474
28. Gibb W. The role of prostaglandins in human parturition. Ann Med. 1998; 30(3):235-241
29. Goureau A, Yerle M, Schmitz A, Riquet J, Milan D, Pinton P, Frelat G, Gellin J.Human and porcine correspondence of chromosome segments using bidirectional chromosome painting. Genomics. 1996; 36(2): 252-262
30. Hagemann A, Nielsen AH, Poulsen K. The uteroplacental renin-angiotensin system: a review. Exp Clin Endocrinol. 1994; 102(3): 252-61
31. Hanna N, Hanna I, Hleb M, et al.. Gestational age - dependent expression of IL- 10 and its receptor in human placental tissues and isolated cytotrophoblasts . J Immunol, 2000 ,164 (11): 5721-5728
32. Johansson M, Ellegren H, Andersson L. Comparative mapping reveals extensive linkage conservation—but with gene order rearrangements—between the pig and the human genomes. Genomics. 1995; 25(3): 682-690
33. King AH, Jiang Z, Gibson JP, Haley CS, Archibald AL. Mapping quantitative trait loci affecting female reproductive traits on porcine chromosome 8.Biol Reprod.2003;68(6):2172-2179
34. Li W, Alfaidy N, Challis JR. Expression of extracellular matrix metalloproteinase inducer in human placenta and fetal membranes at term labor. J Clin Endocrinol Metab. 2004; 89(6): 2897-2904
35. Lindstrom TM, Bennett PR. The role of nuclear factor kappa B in human labour.Reproduction. 2005; 130(5): 569-581
36. Loudon JA, Sooranna SR, Bennett PR, Johnson MR.Mechanical stretch of human uterine smooth muscle cells increases IL-8 mRNA expression and peptide synthesis.Mol Hum Reprod. 2004; 10(12): 895-899
37. Mackler AM, Iezza G, Akin MR, McMillan P, Yellon SM. Macrophage trafficking in the uterus and cervix precedes parturition in the mouse. Biol Reprod. 1999;61(4): 879-883
38. Majzoub JA, Karalis KP. Placental corticotropin-releasing hormone: function and regulation. Am J Obstet Gynecol. 1999; 180(1 Pt 3): S242-6
39. Majzoub JA, Karalis KP.Placental corticotropin-releasing hormone: function and regulation.Am J Obstet Gynecol. 1999;180(1 Pt 3):S242-6
40. Mastorakos G, Ilias I. Maternal and fetal hypothalamic-pituitary-adrenal axes during pregnancy and postpartumAnn N Y Acad Sci. 2003; 997: 136-49
41. McLean M, Bisits A, Davies J, Woods R, Lowry P, Smith R. A placental clock controlling the length of human pregnancy. Nat Med. 1995; 1(5): 460-463
42. Mendelson CR, Condon JC. New insights into the molecular endocrinology of parturition.J Steroid Biochem Mol Biol. 2005; 93(2-5): 113-119
43. Mitchell MD, Romero RJ, Edwin SS, Trautman MS.Prostaglandins and parturition. J Reprod Fertil. 1995; 7(3): 623-632
44. Mohan AR, Sooranna SR, Lindstrom TM, Johnson MR, Bennett PR. The effect of mechanical stretch on cyclooxygenase type 2 expression and activator protein-1 and nuclear factor-kappaB activity in human amnion cells. Endocrinology. 2007;148(4): 1850-1857
45. Moreau L, Charcosset A, Hospital F, Gallais A.Marker-assisted selection efficiency in populations of finite size. Genetics. 1998;148(3):1353-1365
46. Morris RS, Paulson RJ. Ovarian derived prorenin-angiotensin cascade in human reproduction. Fertil Steril. 1994; 62(6): 1105-1114
47. Pieber D, Allport VC, Hills F, Johnson M, Bennett PR. Interactions between progesterone receptor isoforms in myometrial cells in human labour. Mol Hum Reprod. 2001; 7(9): 875-879
48. Poisner AM. The human placental renin-angiotensin system. Front Neuroendocrinol. 1998; 19(3): 232-252
49. Pomini F , Caruso A , Challis JR , et al. . The effects of interleukin - lbeta and tumor necrosis factor - alpha on The activity and expression of prostaglandin H synthase-2 and The NAD~+-dependent 15-hydroxyprostaglandin dehydrogenase in cultured term human Vil - lous trophoblast and chorion trophoblastcells. J Clin Endcrinol Metab , 1999, 84(12): 4645-4651
50. Rauk PN, Chiao JP. Interleukin-1 stimulates human uterine prostaglandin production through induction of cyclooxygenase-2 expression. Am J Reprod Immunol.2000; 43(3): 152-159
51. Rettenberger G, Adham IM, Engel W, Klett C, Hameister H. Assignment of the porcine transition protein TNP1 to chromosome 15q24-q25 by fluorescence in situ hybridization (FISH). Mamm Genome. 1994; 5(4): 249-250
52. Roh CR, Oh WJ, Yoon BK, Lee JH. Up-regulation of matrix metalloproteinase-9 in human myometrium during labour: a cytokine-mediated process in uterine smooth muscle cells.Mol Hum Reprod. 2000; 6(1): 96-102
53. Rohrer GA, Alexander LJ, Hu Z, Smith TP, Keele JW, Beattie CW. A comprehensive map of the porcine genome. Genome Res. 1996; 6(5): 371-391
54. Roth I , Corry DB , Locksley RM, et al. . Human placental cytotrophoblasts produce The immunosuppressive cytokine interleukin - 10 . J Exp Med , 1996,184(2): 539-548
55. Roth I, Corry DB, Locksley RM, Abrams JS, Litton MJ, Fisher SJ. Human placental cytotrophoblasts produce the immunosuppressive cytokine interleukin 10. J Exp Med. 1996; 184(2): 539-48
56. Rothschild MF, Messer LA, Vincent A. Molecular approaches to improved pig fertility. J Reprod Fertil Suppl. 1997; 52: 227-236
57. Sennstrdm MB, Ekman G, Westergren-Thorsson G, Malmstrom A, Bystr(?)m B,Endresen U, MLambo N, Norman M, St(?)bi B, Brauner A. Human cervical ripening, an inflammatory process mediated by cytokines. Mol Hum Reprod.2000; 6(4): 375-381
58. Sooranna SR, Engineer N, Liang Z, Bennett PR, Johnson MR, Imperial College.Parturition Research Group. Stretch and interleukin 1 beta: pro-labour factors with similar mitogen-activated protein kinase effects but differential patterns of transcription factor activation and gene expression. J Cell Physiol.2007; 212(1): 195-206
59. Sooranna SR, Lee Y, Kim LU, Mohan AR, Bennett PR, Johnson MR. Mechanical stretch activates type 2 cyclooxygenase via activator protein-1transcription factor in human myometrial cells.Mol Hum Reprod. 2004; 10(2): 109-113
60. Spotter A, Distl O. Genetic approaches to the improvement of fertility traits in the pig. Vet J. 2006; 172(2): 234-247
61. Stygar D, Wang H, Vladic YS, Ekman G, Eriksson H, Sahlin L.Increased level of matrix metalloproteinases 2 and 9 in the ripening process of the human cervix.Biol Reprod. 2002; 67(3): 889-894
62. Terzidou V, Sooranna SR, Kim LU, Thornton S, Bennett PR, Johnson MR.Mechanical stretch up-regulates the human oxytocin receptor in primary human uterine myocytes.J Clin Endocrinol Metab. 2005; 90(1): 237-246
63. Thomson AJ, Telfer JF, Young A, Campbell S, Stewart CJ, Cameron IT, Greer IA,Norman JE. Leukocytes infiltrate the myometrium during human parturition:further evidence that labour is an inflammatory process. Hum Reprod.1999;14(1):229-236
64.Watari M, Watari H, DiSanto ME, Chacko S, Shi GP, Strauss JF 3rd.Pro-inflammatory cytokines induce expression of matrix-metabolizing enzymes in human cervical smooth muscle cells. Am J Pathol. 1999; 154(6): 1755-1762
65. Weiss A, Goldman S, Shalev E. The matrix metalloproteinases (MMPS) in the decidua and fetal membranes. Front Biosci. 2007; 12: 649-659
66. Wilkie PJ, Paszek AA, Beattie CW, Alexander LJ, Wheeler MB, Schook LB. A genomic scan of porcine reproductive traits reveals possible quantitative trait loci (QTLs) for number of corpora lutea. Mamm Genome. 1999; 10(6): 573-578
67. Xie C, Xu S. Efficiency of multistage marker-assisted selection in the improvement of multiple quantitative traits. Heredity. 1998; 80 (4): 489-498
68. Yellon SM, Mackler AM, Kirby MA. The role of leukocyte traffic and activation in parturition.J Soc Gynecol Investig. 2003; 10(6): 323-338
2.郭万虎,黄既同,丁山河,郭洪峰,廖美福,蒋思文.新清平猪母系选育研究进展.养猪.2005,2:20-21
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16.杨志军.清平猪部分种质特性的研究.[硕士学位论文].武汉:华中农业大学图书馆,1998
17.张付军,李宁,吴常信.动物主基因(QTL)的定位策略.农业生物技术学报,1996,4(2):166-172
18.张沅.家畜育种学.北京:中国农业出版社,2001
19. Binder ND, Laird MR, Faber JJ. Interrelationships between the renin angiotensin system and uteroplacental blood flow—a recent perspective. Reprod Fertil Dev.1995; 7(6): 1437-1442
20. Brown AG, Leite RS, Strauss JF 3rd. Mechanisms underlying "functional"progesterone withdrawal at parturition. Ann N Y Acad Sci. 2004; 1034:36-1049
21. Casey ML, Cox SM, Word RA, MacDonald PC. Cytokines and infection-induced preterm labour. Reprod Fertil Dev. 1990; 2(5): 499-509
22. Chase AJ, Bond M, Crook MF, Newby AC.Role of nuclear factor-kappa B activation in metalloproteinase-1, -3, and -9 secretion by human macrophages in vitro and rabbit foam cells produced in vivo. Arterioscler Thromb Vasc Biol. 2002;22(5): 765-771
23. Christiaens I, Zaragoza DB, Guilbert L, Robertson SA, Mitchell BF, Olson DM.Inflammatory processes in preterm and term parturition. J Reprod Immunol. 2008;79(1): 50-57
24. Christiaens I, Zaragoza DB, Guilbert L, Robertson SA, Mitchell BF, Olson DM.Inflammatory processes in preterm and term parturition. J Reprod Immunol. 2008;79(1): 50-57
25. Condon JC, Hardy DB, Kovaric K, Mendelson CR.Up-regulation of the progesterone receptor (PR)-C isoform in laboring myometrium by activation of nuclear factor-kappaB may contribute to the onset of labor through inhibition of PR function. Mol Endocrinol. 2006; 20(4): 764-775
26. Condon JC, Jeyasuria P, Faust JM, Mendelson CR. Surfactant protein secreted by the maturing mouse fetal lung acts as a hormone that signals the initiation of parturition. Proc Natl Acad Sci U S A. 2004 ; 101(14): 4978-83
27. Cooper AC, Robinson G, Vinson GP, Cheung WT, Broughton Pipkin F. The localization and expression of the renin-angiotensin system in the human placenta throughout pregnancy.Placenta. 1999; 20(5-6): 467-474
28. Gibb W. The role of prostaglandins in human parturition. Ann Med. 1998; 30(3):235-241
29. Goureau A, Yerle M, Schmitz A, Riquet J, Milan D, Pinton P, Frelat G, Gellin J.Human and porcine correspondence of chromosome segments using bidirectional chromosome painting. Genomics. 1996; 36(2): 252-262
30. Hagemann A, Nielsen AH, Poulsen K. The uteroplacental renin-angiotensin system: a review. Exp Clin Endocrinol. 1994; 102(3): 252-61
31. Hanna N, Hanna I, Hleb M, et al.. Gestational age - dependent expression of IL- 10 and its receptor in human placental tissues and isolated cytotrophoblasts . J Immunol, 2000 ,164 (11): 5721-5728
32. Johansson M, Ellegren H, Andersson L. Comparative mapping reveals extensive linkage conservation—but with gene order rearrangements—between the pig and the human genomes. Genomics. 1995; 25(3): 682-690
33. King AH, Jiang Z, Gibson JP, Haley CS, Archibald AL. Mapping quantitative trait loci affecting female reproductive traits on porcine chromosome 8.Biol Reprod.2003;68(6):2172-2179
34. Li W, Alfaidy N, Challis JR. Expression of extracellular matrix metalloproteinase inducer in human placenta and fetal membranes at term labor. J Clin Endocrinol Metab. 2004; 89(6): 2897-2904
35. Lindstrom TM, Bennett PR. The role of nuclear factor kappa B in human labour.Reproduction. 2005; 130(5): 569-581
36. Loudon JA, Sooranna SR, Bennett PR, Johnson MR.Mechanical stretch of human uterine smooth muscle cells increases IL-8 mRNA expression and peptide synthesis.Mol Hum Reprod. 2004; 10(12): 895-899
37. Mackler AM, Iezza G, Akin MR, McMillan P, Yellon SM. Macrophage trafficking in the uterus and cervix precedes parturition in the mouse. Biol Reprod. 1999;61(4): 879-883
38. Majzoub JA, Karalis KP. Placental corticotropin-releasing hormone: function and regulation. Am J Obstet Gynecol. 1999; 180(1 Pt 3): S242-6
39. Majzoub JA, Karalis KP.Placental corticotropin-releasing hormone: function and regulation.Am J Obstet Gynecol. 1999;180(1 Pt 3):S242-6
40. Mastorakos G, Ilias I. Maternal and fetal hypothalamic-pituitary-adrenal axes during pregnancy and postpartumAnn N Y Acad Sci. 2003; 997: 136-49
41. McLean M, Bisits A, Davies J, Woods R, Lowry P, Smith R. A placental clock controlling the length of human pregnancy. Nat Med. 1995; 1(5): 460-463
42. Mendelson CR, Condon JC. New insights into the molecular endocrinology of parturition.J Steroid Biochem Mol Biol. 2005; 93(2-5): 113-119
43. Mitchell MD, Romero RJ, Edwin SS, Trautman MS.Prostaglandins and parturition. J Reprod Fertil. 1995; 7(3): 623-632
44. Mohan AR, Sooranna SR, Lindstrom TM, Johnson MR, Bennett PR. The effect of mechanical stretch on cyclooxygenase type 2 expression and activator protein-1 and nuclear factor-kappaB activity in human amnion cells. Endocrinology. 2007;148(4): 1850-1857
45. Moreau L, Charcosset A, Hospital F, Gallais A.Marker-assisted selection efficiency in populations of finite size. Genetics. 1998;148(3):1353-1365
46. Morris RS, Paulson RJ. Ovarian derived prorenin-angiotensin cascade in human reproduction. Fertil Steril. 1994; 62(6): 1105-1114
47. Pieber D, Allport VC, Hills F, Johnson M, Bennett PR. Interactions between progesterone receptor isoforms in myometrial cells in human labour. Mol Hum Reprod. 2001; 7(9): 875-879
48. Poisner AM. The human placental renin-angiotensin system. Front Neuroendocrinol. 1998; 19(3): 232-252
49. Pomini F , Caruso A , Challis JR , et al. . The effects of interleukin - lbeta and tumor necrosis factor - alpha on The activity and expression of prostaglandin H synthase-2 and The NAD~+-dependent 15-hydroxyprostaglandin dehydrogenase in cultured term human Vil - lous trophoblast and chorion trophoblastcells. J Clin Endcrinol Metab , 1999, 84(12): 4645-4651
50. Rauk PN, Chiao JP. Interleukin-1 stimulates human uterine prostaglandin production through induction of cyclooxygenase-2 expression. Am J Reprod Immunol.2000; 43(3): 152-159
51. Rettenberger G, Adham IM, Engel W, Klett C, Hameister H. Assignment of the porcine transition protein TNP1 to chromosome 15q24-q25 by fluorescence in situ hybridization (FISH). Mamm Genome. 1994; 5(4): 249-250
52. Roh CR, Oh WJ, Yoon BK, Lee JH. Up-regulation of matrix metalloproteinase-9 in human myometrium during labour: a cytokine-mediated process in uterine smooth muscle cells.Mol Hum Reprod. 2000; 6(1): 96-102
53. Rohrer GA, Alexander LJ, Hu Z, Smith TP, Keele JW, Beattie CW. A comprehensive map of the porcine genome. Genome Res. 1996; 6(5): 371-391
54. Roth I , Corry DB , Locksley RM, et al. . Human placental cytotrophoblasts produce The immunosuppressive cytokine interleukin - 10 . J Exp Med , 1996,184(2): 539-548
55. Roth I, Corry DB, Locksley RM, Abrams JS, Litton MJ, Fisher SJ. Human placental cytotrophoblasts produce the immunosuppressive cytokine interleukin 10. J Exp Med. 1996; 184(2): 539-48
56. Rothschild MF, Messer LA, Vincent A. Molecular approaches to improved pig fertility. J Reprod Fertil Suppl. 1997; 52: 227-236
57. Sennstrdm MB, Ekman G, Westergren-Thorsson G, Malmstrom A, Bystr(?)m B,Endresen U, MLambo N, Norman M, St(?)bi B, Brauner A. Human cervical ripening, an inflammatory process mediated by cytokines. Mol Hum Reprod.2000; 6(4): 375-381
58. Sooranna SR, Engineer N, Liang Z, Bennett PR, Johnson MR, Imperial College.Parturition Research Group. Stretch and interleukin 1 beta: pro-labour factors with similar mitogen-activated protein kinase effects but differential patterns of transcription factor activation and gene expression. J Cell Physiol.2007; 212(1): 195-206
59. Sooranna SR, Lee Y, Kim LU, Mohan AR, Bennett PR, Johnson MR. Mechanical stretch activates type 2 cyclooxygenase via activator protein-1transcription factor in human myometrial cells.Mol Hum Reprod. 2004; 10(2): 109-113
60. Spotter A, Distl O. Genetic approaches to the improvement of fertility traits in the pig. Vet J. 2006; 172(2): 234-247
61. Stygar D, Wang H, Vladic YS, Ekman G, Eriksson H, Sahlin L.Increased level of matrix metalloproteinases 2 and 9 in the ripening process of the human cervix.Biol Reprod. 2002; 67(3): 889-894
62. Terzidou V, Sooranna SR, Kim LU, Thornton S, Bennett PR, Johnson MR.Mechanical stretch up-regulates the human oxytocin receptor in primary human uterine myocytes.J Clin Endocrinol Metab. 2005; 90(1): 237-246
63. Thomson AJ, Telfer JF, Young A, Campbell S, Stewart CJ, Cameron IT, Greer IA,Norman JE. Leukocytes infiltrate the myometrium during human parturition:further evidence that labour is an inflammatory process. Hum Reprod.1999;14(1):229-236
64.Watari M, Watari H, DiSanto ME, Chacko S, Shi GP, Strauss JF 3rd.Pro-inflammatory cytokines induce expression of matrix-metabolizing enzymes in human cervical smooth muscle cells. Am J Pathol. 1999; 154(6): 1755-1762
65. Weiss A, Goldman S, Shalev E. The matrix metalloproteinases (MMPS) in the decidua and fetal membranes. Front Biosci. 2007; 12: 649-659
66. Wilkie PJ, Paszek AA, Beattie CW, Alexander LJ, Wheeler MB, Schook LB. A genomic scan of porcine reproductive traits reveals possible quantitative trait loci (QTLs) for number of corpora lutea. Mamm Genome. 1999; 10(6): 573-578
67. Xie C, Xu S. Efficiency of multistage marker-assisted selection in the improvement of multiple quantitative traits. Heredity. 1998; 80 (4): 489-498
68. Yellon SM, Mackler AM, Kirby MA. The role of leukocyte traffic and activation in parturition.J Soc Gynecol Investig. 2003; 10(6): 323-338