摘要
目的研究猪带绦虫重组BCG-TSOL18疫苗的稳定性。方法将猪带绦虫重组质粒pMV261-TSOL18电穿孔转化入卡介苗(Bacillus Calmette-Guerin,BCG),获得猪带绦虫重组BCG-TSOL18菌株,实验设野生型BCG对照,连续培养10d,用分光光度计测定600nm处吸光度A值,绘制生长曲线;人工传代10次,通过PCR分析其稳定性。结果同期培养的野生型BCG与重组BCG-TSOL18第1~7d为对数生长期。培养10d时野生型菌株A600由0.09升至1.24,重组型菌株A600由0.11升至1.28。此后分别进入平台期,且略有下降,其A600分别降至1.12和1.15。重组BCG-TSOL18第1~10代菌落均经PCR检测到TSOL18目的基因,未发生TSOL18基因丢失。结论重组BCGTSOL18对数生长期为前7d,且能稳定遗传至少10代,为猪带绦虫重组BCG疫苗的进一步研究奠定了基础。
Objective To research the stability of a recombinant BCG-TSOL18 vaccine against Taenia solium. Meth- ods The recombinant plasmid pMV261-TSOL18 was transformed into BCG by electroporation to obtain a recombinant BCG-TSOL18 vaccine against T. solium. After 10 days of culturing, the growth curves of strains were generated by de termining the absorbance at 600 nm （A600nm） with a spectrophotometer. After 10 passages of subculturing, the stability of the recombinant BCG-TSOL18 vaccine was determined with PCR. Wild-type BCG served as the control. Results Wild-type BCG and recombinant BCG-TSOL18 were simultaneously cultured, and days 1-7 represented the logarithmic phase. For the wild-type strain, A600 nm increased from 0.09 to 1.24. For the recombinant strain, A600 nm increased from 0.11 to 1.28. The stationary phase began after day 8. For the wild-type strain, A600 nm decreased to 1.12. For the recombinant strain, A600nm decreased to 1.15. The recombinant BCG-TSOL18 was identified with PCR. The target gene TSOL18 was detected from passages 1 to 10, and the TSOL18 gene was not lost. Conclusion Days 1-7 represented the logarithmic phase for growth of the recombinant BCG-TSOL18. The plasmid pMV261-TSOL18 was stable in BCG for at least 10 passages. These results have laid the foundation for the further study of a recombinant BCG-TSOL18 vaccine against T. solium.
出处
《中国病原生物学杂志》
CSCD
北大核心
2017年第2期148-150,共3页
Journal of Pathogen Biology
基金
贵州省科技厅联合基金项目[黔科合LH字(2015)7537号]
