牦牛FMDV持续感染分离株非结构蛋白P2的克隆及基因特征分析

Molecular characterization of non-structural protein P2 of FMDV in yak persistent infection strain

以FMDV持续感染模型动物牦牛的食道/咽部分离物(O/P液)为反转录模板,用1对特异性引物扩增目的cDNA,然后与pMD18-T载体连接并转化JM109菌株,再经重组质粒电泳、PCR和EcoRⅠ酶切鉴定.序列测定和分析结果表明,分离株与China/99的同源性最高达93.4%,而与Akesu/58序列的同源性为90.3%;推导的氨基酸序列同源性分别为97.8%、96.7%.

Using oesophageal-pharyngeal（O/P）fluid that isolated from FMDV Akesu/58 persistent infection model yak as templet to reverse transcript,amplify its P2 tract with a pair of primers. The amplified cDNA products were cloned into pMD18-T vectors and transformed into JM109, the recombinant plasmids were identified by electrophoresis,EcoR Ⅰ cleavage and PCR. The nucleotide sequence of the expected P2 tract was determined by Sanger＇S DNA sequence method,and amino acid sequence was compared with the P2 tracts of the other seven FMDV strains. It was shown that the homology of P2 tract of isolation strain with China/99 and Akesu/58 strains were 93.4 %, 90. 3 % respectively,and deduced animo acid sequence were 97.8 %,96.7 % respectively.