绵羊肺腺瘤病毒NM株前病毒gag基因的克隆与序列分析

Cloning and Sequencing Analysis gag Gene of Jaagsiekte Sheep Retrovirus Inner Mongolia Strain

参照GenBank中已发表的绵羊肺腺瘤病毒(JSRV)的全基因序列,设计合成3对引物,对JSRVNM株的gag基因分3段进行PCR扩增,经琼脂糖凝胶电泳分析,分别呈3条531、888和949bp的特异条带,将其分别克隆入pMD 18T载体中,进行序列测定并拼接序列,得到完整的gag基因序列。分析结果表明,与南非代表株(基因序列号NC 001494)的gag基因序列比较,核苷酸同源性为89 0%,推导出的氨基酸同源性为90%。与美国代表株(基因序列号AF105220)的gag基因序列比较,核苷酸同源性为86 3%,氨基酸同源性为87%。

In order to amplify gag gene of Jaagsiekte sheep retrovirus Inner Mongolia strain, three pairs of primers were designed according to the GenBank sequence . The fragments of gag gene was obtained by polymerase chain reaction(PCR),then the genes were cloned into pMD-18 T vector and identified by PstI, EcoRI and SalI digestion. The nucleotide and amino acid sequences of NM strain gag gene were compared with the counterpart sequences of South Africa strain(NC-001494) and USA strain (AF105220). The nucleotide and amino acid homology of gag gene were 89.0%,90% and 86.3%,87%, respectively.