促卵泡素是动物垂体前叶分泌的重要繁殖调控激素。采用PCR产物直接测序法对河流型和沼泽型水牛、牦牛和大额牛的FSHβ基因外显子3编码区进行群体变异检测,并进行了生物信息学分析。牛科物种FSHβ基因外显子3编码区均为231 bp。在水牛中共发现16个SNP位点,其中异义替换13个,推测4个异义替换对FSH功能有显著影响(subPSEC<|-3)。沼泽型水牛中发现的SNP位点均与河流型水牛共享,但它们在两类水牛中的群体遗传组成不同。在普通牛、牦牛和大额牛中分别确定8,3,1个SNP位点,但只有SNP307是异义替换,普通牛与牦牛共享该位点,推测其对FSH功能无显著影响(subPSEC>|-3)。在山羊中有11个SNP位点,8个为异义替换,推测只有SNP253对FSH功能有显著影响(subPSEC<|-3)。在绵羊中有3个SNP位点,皆为同义替换。确定水牛特有的核苷酸位点4个,山羊特有的位点1个,普通牛、牦牛和大额牛共有的核苷酸位点4个,山羊和绵羊中共有的位点2个,这些位点编码氨基酸的差异可能引起了不同类型牛科物种间FSHβ亚基功能上的差异性。
Follicle stimulating hormone(FSH) has important effects in regulating animal reproductive function. In this paper,the polymorphisms in the coding region of FSHβgene exon 3 of river buffalo,swamp buffalo,yak andgayal were detected by using PCR with direct sequencing. The data obtained in this study and published about bo-vine species were dealt with by employing the methods of bioinformatics. The coding region in FSHβgene exon 3 contained 231 bp in all the species of Bovidae. In buffalo,we identified 16 SNPs,of which 13 were non-synony-mous,and 4 non-synonymous loci were predicted to have significant effect on FSH function(subPSEC ＜-3). The SNP loci detected in swamp buffalo were all shared with river buffalo,but the populationgenetic composition were different in two types of buffalos. There were 8,3 and 1 SNPs in cattle,yak andgayal,respectively,but only SNP307 was non-synonymous which was shared by yak and cattle and was predicted to have no significant effect on FSH function(subPSEC ＞-3). Ingoat,11 SNPs were found,of which 8 SNPs were non-synonymous,but only SNP253 was predicted to have significant effect on FSH function(subPSEC ＜-3). As for sheep,3 SNPs were found,they are non-synonymous. In this paper,there are 4 and 1 specific nucleotide sites identified in buffalo andgoat,respectively. There are 4 shared nucleotide sites identified among cattle,yaks andgayal,and 2 shared nucleotide sites identified be-
[1] Pierce Jg,Parsons T F.glycoprotein hormones:structure and function[J]. Annu Rev Biochem,1981,50:465-495.
[2] Kim K E,Gordon D F,Maurer R A. Nucleotide sequence of the bovinegene for follicle-stimulating hormone beta-subunit[J]. DNA,1988,7(4):227-233.
[3] Dias J A,Cohen B D,Lindau-Shepard B,et al. Molecular,structural,and cellular biology of follitropin and follitropin receptor[J]. Vitam Horm,2002,64:249-322.
[4] Dai L,Zhao Z H,Zhao R,et al. Effects of novel single nu-cleotide polymorphisms of the FSH beta-sub unitgene on semen quality and fertility in bulls[J]. Anim Rep Sci,2009,114:14-22.
[5] Dai L,Zao Y M,Zangg L,et al. Molecular cloning and sequence analysis of follicle-stimulating hormone beta pol-ypeptide precursor cDNA from the bovine pituitarygland
[J].gen Mol Res,2011,3:1504-1513.
[6] de Kretser D M,Buzzard J J,Okuma Y,et al. The role of activin,follistatin and inhibin in testicular physiology[J]. Mol Cell Endocrinol,2004,225:57-64.
[7] Gharib S D,Roy A,Wierman M E,et al. Isolation and characterization of thegene encoding the beta-subunit of rat follicle-stimulating hormone[J]. DNA,1989,8(5): 339-349.
[8] Ai X L,Chen Xg,Ji Q,et al. Study on the production of bovine FSH by biological techniques[J]. Science in Chi-na(Series B),1995,25(6):604-609.
[9] Jamesons J L,Becker C B,Lindell C M,et al. Human fol-licle-stimulating hormone beta-subunitgene encodes mul-tiple messenger ribonucleic acids[J]. Mol Endocrinol,1988,2(9):806-815.
[10] Vincent A L,Tuggle C K,Rothschild M F,et al. The pro-lactin receptorgene is associated with increased litter 2 期 杨秋丽等: 牛科物种 FSHβ 基因外显子 3 编码区遗传多态性及其生物信息学分析 59 size in pig[C]. Proc 6th World Congrgenet Appl Livest Pro Armidale,Australia,1998,27:15-18.
[11] 赵要凤,李宁,肖璐,等. 猪 FSHβ亚基基因结构区逆转座子插入突变及其与猪产仔数关系的研究
[J]. 中国科学:C 辑,1999(2):81-86.
[12] 赵要风,李宁,陈永福,等. 猪 FSH-β 亚基基因 RFLP的研究初报[J]. 畜牧兽医学报,1998,29(1): 23-26.
[13] 王继卿,胡江,罗玉柱.FSH基因CDS区遗传特征及其与小尾寒羊产羔数关系的研究[J]. 中国畜牧杂志,2010,(011):5-9.
[14] 梁琛,储明星,张建海,等. FSHβ基因 PCR-SSCP多态性及其与济宁青山羊高繁殖力关系的研究[J]. 遗 传,2006,28(9):1071-1077.
[15] 陈宝剑,蒋钦杨,毛献宝,等. 沼泽型水牛促卵泡素基因β亚基的克隆与序列分析[J]. 广西农业科学,2009,40(012):1599-1602.
[16] Thompson J D,Higgins Dg,Gibson T J. Clustal W:im-proving the sensitivity of progressive multiple sequence alignment through sequence weighting,position-specificgap penalties and weight matrix choice[J]. Nucleic Acids Res,1994,22:4673-4680.
[17] Yeh F C,Yang R C,Boyle T. Popgene microsoft win-dows-based freeware for populationgenetic analysis re-lease 1. 31[M]. Edmonton:University of Alberta,1999.
[18] Stephens M,Smith N,Donnelly P. A new statistical method for haplotype reconstruction from Population data[J]. American Journal of Humangenetics,2001,68,978-989.
[19] Tamura K,Dudley J,Nei M,et al. MEGA4:molecular ev-olutionarygenetics analysis(MEGA) software version 4. 0[J]. Mol Biol Evol,2007,24(8):1596-1599.
[20] Brunham L R,Singaraja R R,Pape T D,et al. Accurate prediction of the functional significance of single nucleo-tide polymorphisms and mutations in the ABCA1gene
[J]. PLoSgenet,2005,1:83.
[21] 冯富彦,李新建,吕刚,等.豫南黑猪及3个引进猪种FSHβ和 PRLR基因多态性与产仔性能的关系
[J]. 华北农学报,2011,26(5):66-70.
[22] 周俊,虞德兵,洪坤月,等.鸡FSH和EXRα基因SNPs分析与早期产蛋性能关系[J]. 华北农学报,2008,23(5):49-52.
[23] Ishak A B L,Sumantri C,Noor R R,et al. Identification of polymorphism of FSH beta-subunitgene as sperm quality marker in Bali cattle using PCR-RFLP[J]. J In-donesian Trop Anim Agric,2011,36(4):221-227.
