摘要
目的 :裂殖子表面抗原 2基因 (Merozoitesurfaceantigen 2 ,MSA2 )是恶性疟原虫的一种保护性抗原 ,为研究其重组BCG疫苗的保护作用 ,首先探讨携带有恶性疟原虫FCC 1 HN株MSA2基因的重组分枝杆菌-大肠杆菌穿梭质粒pBCG MSA2在卡介苗 (BacillusCalmetteGuerin ,BCG)中的表达情况。方法 :采用电穿孔转化法将重组质粒pBCG MSA2导入BCG中 ,通过卡那霉素抗性筛选并经PCR鉴定的重组BCG培养于Middlebrook 7H9Broth (M7H9)培养基 ,并添加 10 %M7H9EnrichmentADC和 0 0 5 %Tween80 ,4周后于 4 5℃进行诱导表达 ,表达产物进行十二烷基磺酸钠 -聚丙烯酰胺凝胶电泳 (Sodiumdodecylsulfatepolyacrylamidegelelectrophoresis,SDS PAGE)及免疫印迹 (Western blot)分析。结果 :SDS PAGE及Western blot分析结果均显示在约 31kDa的位置上可见明显的蛋白条带 ,并与MSA2基因编码序列推断的分子量相符。结论 :恶性疟原虫裂殖子表面抗原 2可在BCG中表达 。
Objective:The merozoite surface antigen 2(MSA2) was supposed to be a protective antigen of Plasmodium falciparum .In order to construct a recombinant Bacillus Calmette Guerin(BCG) vaccine against Plasmodium falciparum ,the recombinant mycobacteria E\^coli shuttle plasmid pBCG/MSA2 carrying the MSA2 gene of Plasmodium falciparum FCC 1/HN was expressed in BCG.Methods:The recombinant plasmid pBCG/MSA2 was introduced into BCG by electroporation,then the recombinant BCG,selected by kanamycin resistance and identified by PCR,was grown in liquid Middlebrook 7H9 medium supplemented with 10% Middlebrook 7H9 Enrichment ADC and 0\^05% Tween 80(M ADC TW broth)with shaking at 37℃.After 4 wk cultivation,the culture was induced at 45℃,and its expressed products were analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE) and Western blot. Results:The expressed MSA2 band could be observed by SDS-PAGE and Western blot at the position of 31kDa,as the predicted molecular weight deduced from MSA2 gene sequences.Conclusion:The MSA2 gene could be expressed in BCG,this will provide scientific evidences for development of BCG as multivalent vectoral vaccine.
出处
《寄生虫与医学昆虫学报》
CAS
2002年第4期193-197,共5页
Acta Parasitologica et Medica Entomologica Sinica
