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合成编码一种人精子膜蛋白YWK-Ⅱ胞外区的一段多肽片段的双链寡核昔酸链,HSD—2a。用平端连接的方法将其插入到沙门氏菌鞭毛基因fliC(d)的抗原表位IV高变区EcoRV位点,构建了重组质粒pLS408-H1。重组基因在鞭毛负性aroA基因缺失的无致病性沙门氏菌S.dublin SL5928疫苗菌株中表达。经ELISA、电镜免疫胶体金法检测,表明HSD-2a编码的多肽片段成功地在沙门氏菌鞭毛表面表达。融合鞭毛蛋白分子量约为60kD。Western-Blot法检测,提纯的融合鞭毛蛋白与YWK-Ⅱ小鼠抗血清只在60 kD处出现一条结合带。携带重组质粒pLS408—H1的沙门氏菌疫苗菌株极有可能成为抗生育疫苗。
Synthesis of a human sperm membrane protein YWK-Ⅱ extracellular region of a polypeptide fragment of double-stranded oligonucleotide chain, HSD-2a. The recombinant plasmid pLS408-H1 was constructed by blunt-end ligation into the EcoRV site of the antigenic epitope IV of fliC (d) of Salmonella flagella. The recombinant gene is expressed in a non-pathogenic Salmonella S. dublin SL5928 vaccine strain lacking the flagella-negative aroA gene. ELISA and electron microscopy immunogold assay showed that HSD-2a encoded polypeptide fragment was successfully expressed on the surface of Salmonella flagella. Fusion flagellin molecular weight of about 60kD. Western Blot method detection, purification of the fusion flagellin and YWK-Ⅱ mouse antiserum at 60 kD only a band. Salmonella vaccine strains carrying the recombinant plasmid pLS408-H1 are likely to become anti-fertility vaccines.