【摘要】 采用RTPCR技术扩增了TGEV河北分离株的N基因,长度约为1149bp。将该基因片段进行亚克隆重组到pMD18T质粒载体上,连接转化,鉴定后得到阳性重组质粒pTN。对重组质粒插入的目的基因片段进行序列分析和比较,结果表明TGEV河北分离株N基因与国外的Purdue115、FS772/70、TO14、961933株有≥95%的同源性;推导的氨基酸序列同源性≥95%。TGEVN基因的克隆及序列分析,为其蛋白质的表达分析提供了重要依据,也为进一步讨论该病毒的分子生物学特性奠定了理论基础。更多还原

【Abstract】 The viral subgenome mRNA of porcine transmissible gastroenteristis virus(TGEV) isolated from Hebei was amplified with RT-PCR. A DNA fragment about 1 149 bp was amplified. It contained the complete ORF of N gene. PCR products were purified and then cloned into plasmid vector PMD18-T. The recombinant plasmid was designated as pTN and analyzed by endonecleoase digestion from proper inserts. The sequence analysis of the insert fragment in the recombinant pTN indicated that TGEV isolated from Hebei shared more than 95% with Purdue-115, FS772/70, TO14 and 96-1933 strains. The amino acid sequence homology was not less than 95%. The clone and sequence analysis of N gene from TGEV provided a basis for the study of protein expressing and molecular characteristics.更多还原