Related Experiment Videos
[Gene cloning and expression of Mycobacterium leprae alpha 2 antigen]
1Department of Experimental Diagnosis, Institute of Dermatology, CAMS and PUMC, Nanjing 210042.
Objectives:
The recombinant alpha 2 antigen of M. leprae was prepared using the molecular biologic tools and the recombinant DNA expression technology.
Methods:
Screening of the M. leprae expression library was performed by the plaque hybridization technique. Nucleotide sequences were determined by dideoxy termination method.
Results:
The gene coding for alpha 2 antigen of M. leprae was cloned and characterized, and the complete nucleotide sequence data has been assigned in the GSDB, DDBJ, EMBL and NCBI nucleotide sequence databank. The over expression system of alpha 2 antigen gene in E. coli was constructed, and the recombinant alpha 2 antigen has been purified by amylose column chromatography at the purity of more than 95%. More than 10 mg of recombinant alpha 2 antigen has been obtained from 200 ml of liquid culture.
Conclusion:
The recombinant alpha 2 antigen of M. leprae could be used as one of the specific antigens for the sero-diagnosis of leprosy.