Gao A W, Yan Y C, Yang Q S, Zhao F, Dong Q, Zhang M L, Koide S S
Shanghai Institute of Cell Biology, Academia Sinica, Shanghai 200031, China.
Shi Yan Sheng Wu Xue Bao. 1998 Mar;31(1):49-60.
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.
将编码人精子膜蛋白YWK-II细胞外结构域肽段的合成寡核苷酸HSD-2a，通过平端连接插入到沙门氏菌鞭毛蛋白基因fliC（d）高变、抗原决定区IV的EcoRV位点。将重组质粒（pLS408-H1）转入无致病性的aroA活疫苗沙门氏菌都柏林菌株SL5928，该菌株无鞭毛蛋白。使用小鼠抗YWK-II抗血清，通过ELISA和免疫金标记电子显微镜证实了HSD-2a蛋白位于鞭毛表面。从重组都柏林沙门氏菌中分离出一种与嵌合鞭毛蛋白相对应的60 kD蛋白，使用抗YWK-II抗血清通过蛋白质印迹分析发现其含有表达的HSD-2a肽段。因此，携带重组质粒-pLS408-H1的沙门氏菌菌株可能是抗生育疫苗的潜在来源。
