摘要
为了构建Ⅰ型鸭肝炎病毒双拷贝VP1基因的原核表达质粒,试验采用RT-PCR技术扩增鸭肝炎病毒VP1基因,将其连接到克隆载体pGEM-T Easy中得到重组克隆质粒pGEM-T-VP1,阳性克隆质粒经鉴定正确后,分别用BamHⅠ、XholⅠ、BglⅡ进行酶切,重组后得到pGEM-T-2VP1,然后将双拷贝VP1基因亚克隆入原核表达载体pET-30a(+)中,得到重组质粒pET-30a(+)-2VP1。结果表明:得到了含有目的基因的阳性克隆。说明Ⅰ型鸭肝炎病毒双拷贝VP1基因原核表达质粒构建成功。
To construct a recombinant expressing plasmid including two VP1 genes （2VP1） of DHV, according to complete genome of Duck hepatitis virus 1 （ DHV - 1 ）, two special primer pair were designed to amplify VPlgenes （2VP1） of DHV. The gene of VPlwas cloned into the vector T, then it was subcloned into the prokaryotic expression vector pET - 30a（ ＋ ） after the sequence was identified by BamH I , Xho II, Bg III digestion and sequencing analysis. The results showed that the ligation direction of the objective gene fragment was correct. It indicates that the prokaryotic expression vector pET -30a -2VP1 has been constructed successfully.
出处
《黑龙江畜牧兽医》
CAS
北大核心
2011年第8期15-17,共3页
Heilongjiang Animal Science And veterinary Medicine
基金
国家自然科学基金项目(30871877)
关键词
I型鸭肝炎病毒
2VP1基因
串联表达
Duck hepatitis virus （ DHV ） serotype 1 （ DHV - 1 ）
2VP1 gene
tandem expression
