山羊卵泡刺激素α亚基cDNA的分子克隆与序列分析

Nucleotide Sequence of Cloned cDNA for alpha Subunit of Goat Follicle Stimulating Hormone

从新屠宰的雌山羊脑垂体中提取总RNA ,反转录获得cDNA .以此cDNA为模板用PCR法扩增目的片段 ,获得长为 380bp的山羊卵泡刺激素α亚基cDNA片段 .将它克隆至pMD 18 T Verctor.随机挑选 3个阳性重组子进行测序 ,将测序结果与绵羊、牛、猪等多种哺乳动物该基因的核苷酸序列及相应氨基酸序列进行比较 .结果表明 ,山羊卵泡刺激素α亚基基因氨基酸序列与绵羊、水牛的同源性最高 ,达 96 % ,与牛的同源性达 95 % ,与人的同源性较低 ,为 74 % .山羊卵泡刺激素α亚基基因编码区的核苷酸与绵羊的同源性最高 ,达 95 % ,与水牛、牛的同源性达 94 % ,与马和大鼠的同源性较低 ,为 85 % .总体来看 ,在哺乳类动物中FSHα亚基基因同源性还是很高的 .

Total RNA was prepared from pituitary gland of new butchered female goat.cDNA was synthesized by RT PCR and was used as a model in PCR amplification for α subunit of goat follicle stimulating hormone.The PCR product was 380 bp as determined by 1.2 % agarose gel electrophoresis with the same length as FSH α subunit.After cloning it to pMD\|18\|T\|vector and selecting 3 positive recombinants at random the DNA sequences were then analyzed.Comparison of their amino acid sequences with those of other mammalian species showed that goat had the highest homology of 96% with sheep and buffalo with only 5 amino acids difference;and high homology of 95% with bovine;and lower homology of 74% with human.It had the highest homology of 95% with sheep in nucleotide sequence,and high homology of 94% with bovine and buffalo,and lower homology of 85% with horse and rat.It was found that the nucleotide sequence of the FSH α subunit in these mammalian species were highly conservative.Goat FSH α subunit gene was reported.