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Position: Home > Articles > Cloning and Sequencing of Full Vp4 Gene of Group A Porcine Rotavirus Journal of Jilin Agricultural University 2005,27 (5)

猪A组轮状病毒Vp4全基因的克隆与序列分析

作  者:
王春凤;于守平;杨树宝;李玉
单  位:
吉林农业大学菌物研究所;吉林农业大学动物科学技术学院
关键词:
轮状病毒;Vp4全基因;克隆与测序
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。
译  名:
Cloning and Sequencing of Full Vp4 Gene of Group A Porcine Rotavirus
作  者:
WANG Chun-feng~1,YU Shou-ping~1,YANG Shu-bao~1,LI Yu~2(1.College of Animal Science and Technology,Jilin Agricultural University,Changchun 130118,China;2.Institute of Mycology,Jilin Agricultural University,Changchun 130118,China)
关键词:
rotavirus;full Vp4 gene;cloning and sequencing
摘  要:
The antigenic determinants of Vp4 gene of porcine group A Rotavirus were amplified from cell infected with rotavirus 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 rotavirus Vp4 gene has over 99% similarity with the rotavirus Vp4 genes of America strain and Heilongjiang strain,which has a positive open reading frame.

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