The construction of Schistosoma japonicum vaccine BCG-Sj26GST and its identification
| Autor: | Huangfu Yong-mu, Feng Zuohua, Cheng Jizhong, Liang Juqing, Zheng Bo |
|---|---|
| Rok vydání: | 2003 |
| Předmět: |
Recombinant Fusion Proteins
Genetic Vectors Biology Polymerase Chain Reaction Schistosoma japonicum law.invention Microbiology Plasmid law Management of Technology and Innovation Complementary DNA Gene expression Escherichia coli Animals Humans HSP70 Heat-Shock Proteins Cloning Molecular Gene Vaccines Synthetic Mycobacterium tuberculosis biology.organism_classification Molecular biology Antigens Helminth Recombinant DNA BCG Vaccine BCG vaccine Mycobacterium |
| Zdroj: | Journal of Tongji Medical University = Tong ji yi ke da xue xue bao. 19(3) |
| ISSN: | 0257-716X |
| Popis: | The expression of foreign gene, Schistosoma Japonicum 26 ku antigen (Sj26GST), in Bacillus Calmette-Guerin (BCG), Mycobacterium (M. smegmatis) and Escherichia coli (E. coli) were studied. The cDNA fragment encoding Sj26GST was amplified by PCR using plasmid pGEX, which could express Sj26GST in E. coli as template. The Sj26GST cDNA was cloned into the down-stream of human M. tuberculosis heat shock protein (hsp) 70 promoter with correct reading frame, and then the DNA fragment containing hsp70 promoter and Sj26GST gene were subcloned together into E. coli-Mycobacteria shuttle plasmid pBCG-2000 to construct the expression shuttle plasmid pBCG-Sj26. The recombinant BCG and M. smegmatis mc(2)155, which were electroplated with pBCG-Sj26, could express Sj26GST and the recombinant Schistosoma Japonicum vaccine BCG-Sj26GST was made. The recombinant Sj26GST (rSj26GST) were soluble and could be observed on SDS-PAGE at molecular weight of 26 ku. The content of rSj26GST accounted for 15% and 10% of total bacterial protein in BCG and M. smegmatis respectively. The results of Western blot showed the combination of rSj26GST with antibody of GST. |
| Databáze: | OpenAIRE |
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