问号状钩端螺旋体赖型017株OmpL1基因重组穿梭质粒在卡介苗中的构建及表达
The construction and expression of recombinant shuttle plasmid with OmpL1 gene from leptospira interrogans serovar Lai strain 017 in Bacille Calmette-Guerin.
作者信息
Bao Lang, Qiu Hongyu, Yan Jufang, Xie Yongen, Chen Wei
机构信息
Research Unit of Leptospirosis & Infection and Immunity, West China University of Medical Sciences, Chengdu 610041.
出版信息
Chin Med Sci J. 2002 Jun;17(2):81-4.
OBJECTIVE
To construct recombinant BCG against leptospirosis.
METHODS
We amplified the entire open reading frame of the OmpL1 gene from the genome of the leptospire serovar Lai strain 017. Two recombinant plasmids pBQ1 and pBQ2 were constructed by oriented ligation based on the E. coli-BCG shuttle plasmids pMV261 and pMV361 respectively. The recombinant plasmids were transformed into BCG by electroporation. The rBCGs bearing pBQ1 and pBQ2 were induced by high temperature of 45 degrees C.
RESULTS
The expressed product, a 35kD protein was detected by SDS-PAGE. The result indicates that pBQ1 and pBQ2 can express OmpL1 in rBCG.
CONCLUSION
The technical methods in this study may help detect the immunogenicity and immunoprotection of OmpL1 and develop more safe, highly effective rBCG bearing leptospiral antigen with long-lasting protection.
目的
构建抗钩端螺旋体病的重组卡介苗。
方法
从钩端螺旋体赖型菌株017基因组中扩增出OmpL1基因的完整开放阅读框。分别基于大肠杆菌-卡介苗穿梭质粒pMV261和pMV361，通过定向连接构建了两个重组质粒pBQ1和pBQ2。通过电穿孔将重组质粒导入卡介苗。携带pBQ1和pBQ2的重组卡介苗经45℃高温诱导。
结果
通过SDS-PAGE检测到表达产物为一种35kD的蛋白质。结果表明pBQ1和pBQ2可在重组卡介苗中表达OmpL1。
结论
本研究中的技术方法可能有助于检测OmpL1的免疫原性和免疫保护作用，并开发出更安全、高效、具有持久保护作用的携带钩端螺旋体抗原的重组卡介苗。
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