山羊卵泡刺激素α亚基cDNA的分子克隆与序列分析

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 .

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

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.