人精子膜蛋白片段在沙门氏菌中的表达——一种制备抗生育疫苗的途径

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苷酸链，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

A potential approach to antifertility vaccine by expression of sperm membrane peptide in Salmonella.

A synthetic oligonucleotide, HSD-2a, encoding a peptide segment of the extracellular domain of a human sperm membrane protein, YWK-II, was inserted by blunt-end ligation at the EcoRV site in the hypervariable, antigenically determinant region IV of Salmonella flagellin gene fliC (d). The recombinant plasmid (pLS408-H1) was transferred into the nonpathogenic aroA live vaccine Salmonella dublin strain SL5928, which is flagellin-negative. Location of the HSD-2a protein on the surface of the flagella was demonstrated by ELISA and by immunogold-labeling electron microscopy using mouse anti-YWK-II antiserum. A 60 kD protein, corresponding to the chimeric flagellin, was isolated from the recombinant S. dublin and found to contain the expressed HSD-2a peptide determined by Western blot analysis using anti-YWK-II antiserum. The Salmonella strain carrying the recombinant plasmid-pLS408-H1 may thus be a potential source of antifertility vaccine.