猪A组轮状病毒Vp4全基因的克隆与序列分析  

Cloning and Sequencing of Full Vp4 Gene of Group A Porcine Rotavirus

作　　者：王春凤[1] 于守平[1] 杨树宝[1] 李玉[2] 

机构地区：[1]吉林农业大学动物科学技术学院,长春130118 [2]吉林农业大学菌物研究所,长春130118

出　　处：《吉林农业大学学报》2005年第5期549-553,共5页Journal of Jilin Agricultural University

基　　金：国家"863"计划(2003AA241121002);国家自然科学基金(30200199);吉林省科技厅基金(20020220);吉林省杰出青年基金(20030118);中国博士后科学基金(2002031156);吉林农业大学青年教师启动基金(2002-QQN-002)资助项目

摘　　要：应用RT-PCR技术,从猪A组轮状病毒总RNA中扩增了2 331 bp的Vp4全基因,通过T-A克隆技术,将PCR产物克隆至克隆载体pGEM-T中,转化至受体菌株JM109中,通过质粒提取、限制性内切酶酶切、PCR、序列测定等方法鉴定,构建出重组阳性克隆质粒pGEM-T-Vp4。经DNASTAR软件分析核苷酸和氨基酸序列,结果显示该实验株与黑龙江株和美国株的同源性最高,均达到99%以上,并具有轮状病毒Vp4全基因的抗原表位区。The antigenic determinants of Vp4 gene of porcine group A Rotavirus were amplified from cell infected with rotavims by the reverse transcription-polymerase chain reaction （RT- PCR）. The length of full Vp4 gene was 2 331 bp. The products of RT - PCR were ligated with plasmid pGEM - Teasy and transformed to JM109. By the analysis of restriction endonuclease cutting, PCR and sequencing, the recombinant plasmids pGEM - T - Vp4 was constructed. The results of nucleotide and amino acid sequences analysis showed that the experimental rotavims Vp4 gene has over 99% similarity with the rotavirus Vp4 genes of America strain and Heilongjiang strain, which has a positive open reading frame.

关 键 词：轮状病毒 Vp4全基因 克隆与测序 

分 类 号：S852.65[农业科学—基础兽医学]