摘要
目的构建炭疽芽孢杆菌(Bacillus anthracis)A16R株lysA基因缺失突变株,为后续的定量蛋白质组学研究奠定基础。方法以炭疽杆菌活疫苗A16R株lysA基因为目的缺失基因,利用软件设计上下游同源臂以及抗性基因的引物,用同源重组酶将3个片段连入质粒中,构建重组质粒,并将重组质粒导入炭疽杆菌A16R感受态细胞中,筛选炭疽杆菌A16R株lysA基因缺失突变株,对其进行验证。最后绘制缺失突变株和野生株生长曲线并进行生理生化分析。结果成功构建了重组质粒,经同源重组后获得lysA基因缺失突变株。鉴定表明目的基因已经丢失。结论成功获得炭疽杆菌A16R株lysA基因缺失突变株,为定量蛋白质组学研究奠定了基础,也为炭疽杆菌重要基因功能的研究建立了良好的技术平台。
Objective To construct the lysA site-deleted mutagenesis of Bacillus anthracis vaccine strain A16R in order to provide scientific reference for subsequent study on quantitative proteomics. Methods Using lysA Site-deleted mutagenesis as the target gene, software was used to design primers of upstream and downstream of lysA and antibiotic resistance genes. The recombinant plasmid was constructed by inserting three fragments into the vector and electroporated into compe- tence A16R cells. Finally, A16R mutagenesis strain was screened and verified. Growth curves of the mutagenesis strain and wild strain were drawn, and physiological and biochemical characteristics were analyzed. Result and Conclusion lysA Site-deleted mutagenesis is obtained ,contributing to quantitative proteomics research and establishing a good technical plat- form for functional genomics research of B. anthracis.
出处
《军事医学》
CAS
CSCD
北大核心
2013年第2期97-101,113,共6页
Military Medical Sciences
基金
国家自然科学基金资助项目(81071322)
国家科技重大专项资助项目(2012ZX10004215)
关键词
炭疽芽孢杆菌A16R株
同源重组
赖氨酸营养缺陷型
Bacillus anthracis vaccine strain A16R
homologous recombination
lysine auxotrophic mutant
