龋齿DNA疫苗pGJA—P／VAX1生产用菌种库的建立和检定  

Establishment and quality control of Escherichia coil cell bank for producing anti-caries DNA vaccine pGJA-P/VAX1

作　　者：罗丹[1] 闭兰[1] 赵亚杰[1] 张爱华[1] 杨晓明[1] 

机构地区：[1]武汉生物制品研究所有限责任公司免疫学研究室,430060

出　　处：《国际生物制品学杂志》2011年第6期281-284,共4页International Journal of Biologicals

基　　金：国家科技支撑计划课题研究专项（2007BA12804）

摘　　要：目的建立用于龋齿DNA疫苗pGJA—P／VAX1制备的菌种库，并对建立的菌种库进行质量检定。方法将质粒pGJA-P／VAX1转化大肠杆菌DH-Sct，筛选后建立原始菌种库，扩大培养建立主菌种库和工作菌种库。对主菌种库和工作菌种库进行全面检定，包括革兰染色、生化试验、16SrRNA测序、抗生素抗性检测、外源噬菌体检测、质粒传代稳定性检测、质粒酶切分析。结果种子菌的革兰染色、生化试验和16SrRNA测序结果均符合大肠杆菌检定要求，菌种库无噬菌体污染，质粒酶切结果与质粒构建图谱一致。结论主菌种库和工作菌种库的检测结果均符合国家食品药品监督管理局相关指南要求，能用作龋齿DNA疫苗pGJA—P／VAX1的生产用菌种。Objective To establish Escherichia coli （E. coli） cell banks for producing anti-caries DNA vaccine pGJA-P/VAX1, and perform quality control on the established cell banks. Methods pGJA-P/ VAX1 was transformed into E. coli DH-5α, clones with high pGJA-P gene expression were selected, and then the primary cell bank was established. The master and working cell banks were established by magnification culture, and then control tests, including Gram staining, biochemical tests, 16S rRNA gene sequencing, antibiotic resistance examination, exogenous phage determination, plasmid passage stability determination and restriction endonuclease digestion analysis, were carried out on cells in the cell banks. Results The results of Gram staining, biochemical tests and 16S rRNA gene sequencing were all in accord with E. coli characters. No phage contamination was found in cell banks. The pattern between restriction endonuclease digestion and plasmid construction was similar. Conclusions The master and working cell banks meet the requirement of State Food and Drug Administration guideline for DNA vaccine, and have the potential to be the strain for production.

关 键 词：疫苗 DNA pGJA-P/VAX1 菌种库 检定 

分 类 号：R7[医药卫生—临床医学]