摘要
采用RT-PCR技术对猪瘟病毒(CSFV)E2基因进行了克隆、测序和特征分析。结果,得到了长度为1100bp、编码367个氨基酸残基的目的基因片段。将克隆到的E2基因插入到pAdTrack-CMV穿梭质粒中,再将重组穿梭质粒pAdTrack-CMV-E2用PmeⅠ线性化后电转化携带有腺病毒骨架载体pAdEasy-1的大肠杆菌BJ5183感受态细胞,经细菌内同源重组成功获得了含有CSFVE2基因的重组腺病毒表达载体。
A full length E2 gene of classical swine fever virus（CSFV） was amplified and cloned by RT- PCR. The results showed that the E2 gene contained 1 100 bp,encoding 367 amino acids. Then,it was subcloned into the adenovirus shuttle vector pAdTrack-CMV,and the transfer vector pAdTrack-CMV-E2 was constructed. After being linearized, the pAdTrack-CMV-E2 was transformed into the Escherichia coli strain BJ5183 competent cells. The homologous recombination between the linearized genome and the supereoiled adenoviral vector pAdEasy-1 occurred in it,and then the recombinant adenovirus pAdEasy-E2 was collected at the appearance of cytopathic effect,and then purified using the plaque test.
出处
《中国兽医科学》
CAS
CSCD
北大核心
2008年第9期782-786,共5页
Chinese Veterinary Science
关键词
猪瘟病毒
E2基因
腺病毒
表达载体
classical swine fever virus
E2 gene
adenovirus
expression vector
