钩端螺旋体膜表面蛋白LipL41基因的克隆、序列分析及表达

目的:进一步分析钩端螺旋体LipL41的免疫原性.方法:通过PCR的方法,以我国特有的钩端螺旋体流感伤寒群临海型lin6株(56609)的DNA 为模板,扩增目的基因LipL41,克隆至pcDNA3载体上,以自动测序仪测序后进行序列分析,然后克隆至原核表达载体pGEX-5T进行原核表达,Western印迹分析其免疫原性.结果:获得长1 068 bp的片段,DNA序列分析表明该菌株的LipL41基因与文献报道具有很高的同源性(95.0％～99.4％),在大肠杆菌中获得了表达,并与抗钩端螺旋体血清反应.结论:该抗原为致病性钩端螺旋体所具有的保守性抗原成分,可能在钩端螺旋体病的诊断和预防中发挥作用.

Cloning,sequencing and expression of gene encoding surface-exposed lipoprtein lipl41 of leptospira species

Objective: To further analyze the immunogenicity of LipL41, an important surface-exposed lipoprotein onthe outer membrane of Leptospira species. Methods: The LipL41 gene was amplified by PCR from Leptospira serogroup grip-potyphosa, serovar Linhai isolate lin6 (56609), which was one of the characteristic Leptospira species in China, and wascloned into plasmid pcDNA3 then sequence analysis was done. Later, LipL41 was expressed in E. coli K802 by inserting theLipL41 into pGEX 5T, and identified by Western blot analysis. Results: A fragment of 1 068 bp was amplified, and sequenceanalysis showed it had a high homology with those of other pathogenic Leptospira species, fusion protein GST-LipL41 wassuccessfully expressed in E. coli, and reacted with antiaileptospiral serum. Conclusion: It is indicated that LipI-4l is a con-served antigen among the pathogenic Leptospira species, fusion protein GST--LipI-4l may play an important role in diagnosisand prevention of leptospirosis.