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[Cloning and expression of Cysticercus cellulosae antigen cC1 in E. coli]
1Department of Medical Genetics, Second Military Medical University, Shanghai 200433.
Objective:
To clone and express Cysticercus Cellulosae antigen cC1 in E. coli.
Methods:
cC1 cDNA fragment was cloned to BamHI and PstI sites of pGEM-3Z vector. After alteration of the restriction sites, the fragment was cloned to EcoRI and XhoI sites of pGEX-5T with a synthetic linker to construct recombinant expression vector pGEX-5T-cC1.
Results:
The clone produced the largest yield of cC1 protein expression when incubated in 2YT culture medium for 3 h or induced by IPTG for 6 h. Detected by scanning optical densitometry, cC1 constituted 57% of the total bacterial proteins. Western blotting analysis revealed that the GST-cC1 fusion protein exhibited a specific reactive band.
Conclusion:
High level expression of Cysticercus cellulosae antigen cC1 was obtained in E. coli.