[麻风分枝杆菌α2抗原的基因克隆与表达]
[Gene cloning and expression of Mycobacterium leprae alpha 2 antigen].
作者信息
Yin Y, Suzuki Y, Makino M, Wu Q
机构信息
Department of Experimental Diagnosis, Institute of Dermatology, CAMS and PUMC, Nanjing 210042.
出版信息
Zhongguo Yi Xue Ke Xue Yuan Xue Bao. 1999 Feb;21(1):62-8.
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.
目的
利用分子生物学工具和重组DNA表达技术制备麻风杆菌重组α2抗原。
方法
采用噬菌斑杂交技术筛选麻风杆菌表达文库。通过双脱氧末端终止法测定核苷酸序列。
结果
克隆并鉴定了编码麻风杆菌α2抗原的基因，其完整核苷酸序列数据已提交至GSDB、DDBJ、EMBL和NCBI核苷酸序列数据库。构建了α2抗原基因在大肠杆菌中的过表达系统，通过直链淀粉柱层析纯化重组α2抗原，纯度超过95%。从200ml液体培养物中获得了超过10mg的重组α2抗原。
结论
麻风杆菌重组α2抗原可作为麻风病血清学诊断的特异性抗原之一。
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