摘要
目的构建携带炭疽芽孢杆菌保护性抗原(PA)的两种穿梭载体与上游强启动子。方法将解淀粉芽孢杆菌的α-淀粉酶启动子克隆到载体pbluescript-sk(+)上,构建载体pBLKSP;以A16R疫苗株(Tox+,Cap-,弱毒株)DNA为模板,设计合成内外侧引物巢式PCR扩增获得保护性抗原(PA)的全基因,先克隆到载体pBLKSP,再将其和质粒PUB110重组构建成两种穿梭载体。结果酶切鉴定显示所切下的片段大小均与预计相符。测序结果与文献报道序列及预计结果一致。结论成功构建了带有强启动子的两种穿梭载体。为在无毒炭疽疫苗株中的高效表达和炭疽芽孢杆菌的分子疫苗研究奠定了基础。
Objective To construct two shuttle vector of anthrax toxin proteetive antigen with upstream strong promoter. Methods The promoter region of the a - amylase gene of Bacillus amyloliquefaciens was cloned into pblueseript - sk（ ＋ ）vector; the anthrax toxin protective antigen whole gene was amplified by nested PCR from A16 vaccine strain template. Firstly it was cloned into pBLKSP vector, then reeombinated with PUB110 vector to form two kinds of shuttle vectors. Results Restriction analysis and DNA sequencing confirmed that the cloned gene fragments encoded anthrax toxin proteetive antigen. Conclusion Two kinds of shuttle vectors have been suceessfttlly constructed, it has set foundation for further study on the highly effective expression in nontoxic anthraeis strain and molecule vaccine research of Bacillus anthracis.
出处
《中国热带医学》
CAS
2006年第9期1546-1548,共3页
China Tropical Medicine
关键词
炭疽芽孢杆菌
穿梭载体
重组
基因
Bacillus anthracis
Shuttle vector
Recombination
Gene
