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根据钩体内鞭毛蛋白（Ｅｎｄｏｆｌａｇｅｌｌｉｎ）ｆｌａＢ基因核苷酸序列自行设计一对引物，以赖型钩体ＤＮＡ为模板，用ＰＣＲ扩增到约８４０ｂｐ的片段。扩增片段经酶切（ＢａｍＨＩ、ＰｓｔＩ）定向克隆入ｐＵＣ８。重组质粒ｐＬＦ１插入片段与哈勒焦型钩体ｆｌａβ基因相比，核苷酸序列和推测的氨基酸序列同源性很高。ＳＤＳ－ＰＡＧＥ表明有一约３３ｋｄ的特异蛋白带，表达量占菌体蛋白１１％。免疫印迹试验表明此区带能被抗赖型钧体内鞭毛（轴丝）抗血清识别。首次以ＢＡＬＢ／ｃ小鼠钧体病模型为基础，用含重组质粒ｐＬＦ１的大肠杆菌作免疫原进行免疫保护试验，实验组存活率高子对照组，表现了一定程度的保护作用。本研究结果在所查国内外文献中尚属首次报道。
A pair of primers was designed according to the flaB gene nucleotide sequence of the endoflagellin. A fragment of about 840bp was amplified by PCR using the leptospira DNA as a template. The amplified fragment was cloned into pUC8 by restriction enzyme digestion (BamHI, PstI). The nucleotide and deduced amino acid sequence homology of recombinant plasmid pLF1 was higher than that of flaβ gene. SDS-PAGE showed a specific protein band of about 33kd, the expression amount of bacterial protein 11%. Immunoblotting showed that this region was recognized by antiserum against flagella (axonal filaments). For the first time, based on the BALB / c mice model, the E. coli with the recombinant plasmid pLF1 was used as an immunogen for immuno-protection test. The survival rate of the experimental group was higher than that of the control group, which showed a certain degree of protective effect. The results of this study are the first reported in the literature examined.