杂交牛(大额牛×云南黄牛)朊蛋白基因 的分子克隆及其序列分析

朊蛋白(prion protein,PRNP)基因编码朊蛋白,是引起疯牛病的主效基因。本研究利用PCR方法首次从杂交牛(大额牛×云南黄牛)基因组中扩增了PRNP基因,GenBank登录号为HQ875337。PCR产物直接双向测序表明,该序列包含杂交牛PRNP基因795 bp的开放阅读框(ORF),编码264个氨基酸前体蛋白。生物信息学分析结果发现,该蛋白包含1个信号肽、3个α螺旋、2个β折叠、6个八肽重复序列、1个疏水区域、1个二硫键和1个糖基磷脂酰肌醇锚定位点。与已报道的其他牛PRNP基因进行序列比对分析,核苷酸和氨基酸的同源性均在97%以上。

Molecular Cloning and Sequences Characteristics Analysis of the Prion Protein Gene from the Hybrids between Gayal(Bos frontalis) and Yunnan Yellow Cattle(Bos taurus)

The prion protein was encoded by the prion protein(PRNP) gene which was the major gene for affecting mad cow disease or BSE.In the present study,the PCR method was used to amplify the PRNP gene from the hybrids between Gayal(Bos frontalis) and Yunnan Yellow cattle(Bos taurus).The sequence was deposited in the GenBank under accession number HQ875337.The PCR products were sequenced bi-directly.By sequence analysis,the length of open read frame(ORF) of the PRNP gene was 795 bp,which encodes a protein of 264 amino acids,including a signal region,a octapeptide repeat,three α-helices,two β-sheets,a hydrophobic region,a disulfide bridge,a glycosyl phosphatidyl inositol(GPI) anchor site.The homology of both nucleotide and amino acid sequences with other cattle was more than 97%.All this will provide the molecular basal data for breakthrough of transmissible spongiform encephalopathy among different animal species.