猪带绦虫TSOL18基因重组乳球菌疫苗的稳定性分析  

Stability of the recombinant Lactococcus lactis vaccine of Taenia solium TSOL18 gene

作　　者：周必英[1] 孙俊超[1] ZHOU Bi-ying;SUN Jun-chao(Department of Parasitology,Zunyi Medical College,Zunyi 563000,China)

机构地区：[1]遵义医学院寄生虫学教研室,遵义563000

出　　处：《中国人兽共患病学报》2018年第11期1016-1020,共5页Chinese Journal of Zoonoses

基　　金：贵州省优秀青年科技人才基金项目(No.黔科合平台人才[2017]5629);贵州省卫计委科研项目(No.gzwjkj2016-1-048);省市科技合作专项资金项目(省市科合[2015]52)~~

摘　　要：目的研究猪带绦虫TSOL18基因重组乳球菌(Lactococcus lactis,L.lactis)疫苗的人工传代稳定性。方法将重组质粒pMG36e-TSOL18、pMG36e-SP-TSOL18电转入L.lactis,经PCR鉴定阳性菌株pMG36e-TSOL18/L.lactis、pMG36e-SP-TSOL18/L.lactis,在无红霉素抗性和含红霉素抗性条件下,连续人工传代20次,通过测定质粒遗传稳定率和双酶切鉴定,分析质粒pMG36e-TSOL18、pMG36e-SP-TSOL18在L.lactis中的稳定性情况。结果质粒pMG36e-TSOL18、pMG36e-SP-TSOL18在L.lactis中连续传代20代后,在无红霉素抗性条件下,质粒稳定率均为100%,在含红霉素抗性条件下,质粒稳定率均为99%。将各代次的质粒采用限制性内切酶SacI/HindIII进行双酶切鉴定,琼脂糖凝胶电泳显示大小正确,连续传至20代时,与原代质粒相符。结论提示质粒pMG36e-TSOL18、pMG36e-SP-TSOL18在L.lactis中连续传代20代时,均有较好的遗传稳定性,为猪带绦虫TSOL18基因重组乳球菌疫苗的进一步研究奠定了基础。We analyzed the stability of the recombinant Lactococcus lactis(L.lactis)vaccine of Taenia solium TSO L18 gene after the artificial generation.These two plasmids of pMG36e-TSO L18 and pMG36e-SP-TSO L18 were electroporated into L.lactis MG1363.Positive clones were identified by PCR.In the absence of erythromycin resistance and erythromycin resistance under the condition of continuous artificial passage for 20 times,the stability of pMG36e-TSO L18 and pMG36e-SP-TSO L18 in L.lactis was analyzed through the determination of plasmid genetic stability rate and Sac I/Hin dIII enzyme digestion.After pMG36e-TSO L18 and pMG36e-SP-TSO L18 were continuously passaged for 20 generations in L.lactis,the stability rate of these two plasmids was 100%in the absence of erythromycin resistance,and the stability rate was 99%under the erythromycin resistance.These plasmids of all generations were identified by restriction endonuclease Sac I/Hin dIII.The agarose gel electrophoresis showed that the size was correct,and it was consistent with the original plasmid when it was continuously transmitted to the 20th generation.It was indicated that pMG36e-TSO L18 and pMG36e-SP-TSO L18 plasmid had good genetic stability when they were continuously passaged for 20 generations in L.lactis,which laid the foundation for further research on TSO L18 gene recombinant L.lactis vaccine of Taenia solium.

关 键 词：猪带绦虫 TSO L18基因 重组乳球菌疫苗 稳定性 

分 类 号：R383.3[医药卫生—医学寄生虫学]