【摘要】 参照GenBank上已发表的内源性绵羊肺腺瘤病毒env基因与5′长末端重复序列保守区序列设计引物,应用PCR技术成功扩增出内源性绵羊肺腺瘤env基因与5′长末端重复序列,将纯化的扩增片段连接pMD18-T克隆载体进行测序获得目的基因序列。应用MEGA4.1、DNASTAR进行序列分析,分析结果表明,内源性env基因具有完整的开放阅读框,内源性env基因序列与内源性绵羊肺腺瘤(AF153615.1)基因同源性高达99.7%,与外源性绵羊肺腺瘤(M80216.1)基因的同源性仅为90.6%;由内源性env基因推导的氨基酸在NCBI进行BLAST比较:与AF153615.1的氨基酸同源性高达100%。内源性env基因编码表达的蛋白具有亲水性。这些结果为研究5′长末端序列对内源性env基因调控等问题提供了重要参考。更多还原

【Abstract】 According to the conservative subsequences of env gene and long terminal repeat(LTR) of endogenous sheep jaagsiekte retrovirus published in GenBank,env gene and LTR gene of endogenous sheep jaagsiekte retrovirus were amplified by PCR.After purification,the fragment was cloned to pMD18-T vector and sequenced.The sequences were analyzed with MEGA4.1 and DNASTAR software.The results showed that the env gene had complete open reading frame,and it had 99.7% nucleotide acid homology to that of endogenous sheep jaagsiekte retrovirus(AF153615.1) but only 90.6% nucleotide acid homology to that of exogenous sheep jaagsiekte retrovirus(M80216.1).The deduced amino acid sequence of the endogenous env gene was compared with those included by NCBI using BLAST software.The results showed that it had 100% amino acid homology to that of endogenous sheep jaagsiekte retrovirus(AF153615.1),and the encoded protein had hydrophilicity.This study provides important reference for studying the regulatory effects of 5′ LTR on endogenous env gene.更多还原