摘要
通过提取尤凯帕病毒 (Yucaipavirus ,PMV 2 )的RNA ,进行RT PCR ,获得了HN基因。序列分析结果表明 ,HN基因的ORF全长为 174 3nt,编码一由 5 80个氨基酸组成的蛋白。HN基因经双酶切后连接入 pTYB 2原核表达载体。酶切鉴定筛选阳性克隆 ,用IPTG诱导表达 ,经SDS PAGE电泳和Western blotting检测均可见一约 110ku的目的条带 ,说明大肠埃希氏菌表达的HN蛋白能与抗PMV 2阳性血清特异性结合。
Yucaipa virus RNA was extracted and the HN gene of the virus was amplified by RT-PCR. Analysis of the HN geme sequence showed that the ORF of the HN gene was composed of 1743 nt encoding a protein with 580 amino acids. After the HN gene was connected with PTYB-2 vector, the recombinant plasmid pTYB-HN was digested by EcoRⅠ and SmaⅠ. The HN gene was transfered into Escherichia coli ER2566. Positive clones were selected through restrict enzyme digestion. After the HN gene was expressed into ER2566 by means of inducing the positive clones with IPTG, an anticipated band was (obtained) by SDS-PAGE and Western-blotting.
出处
《中国兽医科技》
CSCD
北大核心
2004年第3期8-11,共4页
Chinese Journal of Veterinary Science and Technology
