问号赖型钩端螺旋体重组质粒pDL121的初步研究

目的　探讨问号赖型钩端螺旋体抗原基因的特点。方法　应用６ 种常见内切酶（ＢａｍＨＩ，Ｂｇｌ Ⅱ，ＥｃｏＲⅠ，Ｈｉｎｄ Ⅲ，ＰｓｔⅠ，Ｐｖｕ Ⅱ）对从问号赖型钩端螺旋体０１７ 株的基因文库中筛选出来的重组质粒ｐＤＬ１２１ 外源基因进行酶谱分析，同时用Ｄｉｇｏｘｉｎ 标记的ｐＤＬ１２１ 外源基因片段作探针对不同种属的致病性钩端螺旋体和非致病性钩端螺旋体基因进行杂交分析。结果　显示问号赖型钩端螺旋体重组质粒ｐＤＬ１２１ 外源基因没有６ 种内切酶的酶切位点，重组探针与致病性钩体（ｓｅｒｏｖａｒｌａｉｓｔｒａｉｎ０１７，ｓｅｒｏｖａｒ ｈｅｂｄｏｍａｄｉｓｓｔｒａｉｎ ５６６１０ ，ｓｅｒｏｖａｒ ｐｏｍｏｎａｓｔｒａｉｎ ５６６０８）有杂交信号，与非致病性钩体（ｓｅｒｏｖａｒ ｐａｔｏｃ ｓｔｒａｉｎ Ｐａｔｏｃ Ⅰ，ｓｅｒｏｖａｒｉｌｌｉｎｉｓｔｒａｉｎ ３０５５）无杂交信号，亦不识别大肠杆菌。结论　重组质粒ｐＤＬ１２１ 外源基因可能是问号赖型钩端螺旋体的一个新基因，进一步测序分析将揭示该基因本质，同时因该重组探针能鉴别致病性钩体和非致病性钩体，提示其可用于钩端螺旋体病的早期诊断及钩端螺旋体的鉴定和分类。

PRELIMINARY STUDY ON RECOMBINANT PLASMID pDL121 OF LEPTOSPIRA INTERROGANS SEROVAR LAI

Aim:Recombinant plasmid pDL121 of L interrogans serovar lai was analysed Methods:pDL121 was analysed by using 6 different restriction endonucleases and dot blotting Results:The results showed that the restriction map of pDL121 was quite different from other leptospiral genes reported and digoxin labeled recombinant DNA probe of pDL121 could detect the pathogenic leptospires and could not detect the nonpathogenic leptospires Conclusion:The results indicated that the inserted fragment of pDL121 could be a new gene and further research should be done