猪囊虫病疫苗生产用工程菌菌种的稳定性观察

OBSERVATION ON THE STABLITY OF THE GENETIC ENGINEERING BACTERIA USED TO PRODUCTION OF THE VACCINE AGAINST SWINE CYSTICERCOSIS

目的 探讨猪囊虫病疫苗生产用工程菌菌种在传代过程中及不同保存条件下的稳定性。方法 对工程菌菌种按规程进行了 5 2代传代及不同条件下保存 ,检测菌种的质粒、表达量、免疫学活性等方面的变化。结果 经 5 2代传代 ,工程菌及其菌落形态结构及质粒DNA电泳与原始菌种一致 ,SDS -PAGE电泳显示分子量为 6 0kD ,其表达量约占菌体总蛋白的 19%～ 2 4% ,免疫学活性为 4 0× 10 3 ～ 1 6× 10 4 /mg ,与原代菌种基本一致。菌种在 4℃、 - 2 0℃、 - 30℃、 - 70℃下保存 ,菌种稳定性不一致 ,- 70℃最稳定 ,- 2 0℃最差。结论 猪囊虫病基因工程疫苗生产用工程菌菌种经 5 2次人工传代和- 70℃保存 ,其各种特性是稳定的。

Aim To detect the stablity of the genetic engineering bacteria used to produce the vaccine against swine cysticercosis in the course of propagation and in the different preserved conditions Methods The changes of the morphologic structure,recombinant plasmid,expression level and immunie activity were detected during 52 propagations of the stock cultures or in the different preserved condition Result After 52 propagations,the morphologic structure of the bacteria and its colonies,and DNA etectrophoresis of the plasmid were identical to the original cultures SDS-PAGE revealed that the molecular weight of the recombinant protein was 60kD,expression level was 19 to 24% of the whole bacteria,and its immune ctivity was 4 0×10 3 to 1 6×10 4 every micmgmme protein,these were also identical to the stock cultures The stock bacteria were preserved at 4℃,-20℃,-30℃ and -70℃,respectively,their stability was different They were most stable at -70℃ and worst at -20℃.Conclusion The various characters of the recombinant bacteria were stable after 52 propagation and preservation for 15 months at -70℃