贵州黑山羊肌肉生长抑制素基因cDNA克隆及其蛋白抗原表位预测

为通过原核、真核外源表达贵州黑山羊肌肉生长抑制素(MSTN)蛋白,制备MSTN受体拮抗剂或免疫诱导山羊产生MSTN抗体,从而消除MSTN对肌肉生长的抑制作用奠定基础,采用RT-PCR技术扩增贵州黑山羊生长抑制素基因cDNA序列;应用生物信息学相关软件和方法对生长抑制素蛋白二级结构和抗原表位进行预测。结果表明:贵州黑山羊生长抑制素基因编码区全长1 128bp,编码375个氨基酸。生长抑制素蛋白二级结构以无规卷曲为主,有少量的α螺旋和β片层,少见β转角;推测生长抑制素蛋白在二级结构上可能有3个优势抗原表位区域,分别为72～78、235～240和251～254位肽段。

cDNA Cloning and Prediction of Protein Epitope of Myostatin Gene in Guizhou Black Goat

For the further study of expressing the foreign myostatin(MSTN) gene of Guizhou Black Goats in prokaryotic and eukaryotic system,preparing MSTN receptor agonists or inducing MSTN antibody through vaccination in the goats,which can finally lay the foundation of eliminating the muscle growth inhibition of MSTN,the MSTN gene cDNA sequence of Guizhou Black Goats was amplified by RT-PCR,and the MSTN protein secondary structure and epitopes were predicted by bioinformatical software and methods.The results show that the whole MSTN gene encoding sequence consists of 1 128 bp,encoding 375 amino acid.The protein secondary structure of MSTN includes main random coils,a small amount of α-helix or β-sheet,and rare β-turn,where the No.72~78,235~240,and 251~254 peptide segments were supposed to be the three predominant epitopes.