中国人兽共患病杂志2000，Vol.16Issue(4)：9-11,3.
莱姆病螺旋体P39蛋白的基因克隆及在大肠杆菌中的高效表达
MOLECULAR CLONING AND EXPRESSION IN BORRELIA BURGDORFERI P39 GENE E. COLI
佟玉品 1冯方波 1毕胜利 2江永珍 2鲁健2
作者信息
- 1. 北京军区第二六一医院检验科北京，100094
- 2. 中国预防医科院病毒所
- 折叠
摘要
Abstract
Aim Molecular cloning and expressing BmpA(P39) gene from a Borrelia burgdorferi strain isolated from China for preparing recombinant antigen to the basic and chinical diagnostic study on Lyme disease in China. Method PCR and gene recombination technique were used to clone the whole P39 gene from a strain BT01. The P39 gene was then inserted into a expression vector LKB2 without its signal peptide and expressed in E. coli. The expressed protein was identified by SDS -PAGE,Western blot, and scanning analysis. Results Sequence analysis showed the P39 gene was 1020 base pairs with high homology to foreign isolates. Especially it had 99％ nucleotide and amino acid identity with the Sh-2-82 strain. The recombinant plasmid BT-p39 expressed the protein with high level and a native form in the host cell BL21 (DE3),the protein was about 37KDa,and it accounted for 58％ of total bacterial protein by scanning analysis. Westernblot assay further proved it had good antigenicty to the specific antibody from patients with Lyme disease. Conclusion The P39 gene of the Chinese Borrelia burgdorferi isolate was successfully cloned and highly expressed in prokaryotic expression system. The recombinant P39 protein may provide a promising candidate antigen for diagnostic usage in Lyme disease.关键词
伯氏疏螺旋体 BmpA基因 基因表达 莱姆病Key words
Borrelia burgdorferi/BmpA gene/Gene Expression/Lyme disease分类
医药卫生引用本文复制引用
佟玉品,冯方波,毕胜利,江永珍,鲁健..莱姆病螺旋体P39蛋白的基因克隆及在大肠杆菌中的高效表达[J].中国人兽共患病杂志,2000,16(4):9-11,3.
