Kuang Y, Yan Y C, Gao A W, Zhai Y M, Miao S Y, Wang L F, Koide S S
Institute of Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences.
Arch Androl. 2000 Nov-Dec;45(3):169-80. doi: 10.1080/01485010050193940.
The HSD-I gene codes a human sperm membrane protein (hSMP-1) and has been assigned the accession number U12978. The gene is located on human chromosome 9, region p12-p13. When the 1.7-kb cDNA of HSD-I was digested sequentially with EcoRI, BamHI, and HindIII, a 550-bp cDNA fragment was formed, which codes for the extracellular domain. This fragment was cloned into the asd+ vector pYA3149 to construct pYA3149R. The recombinant plasmid was used to transform an avirulent deltacva, deltacrp, deltaasd vaccine strain of Salmonella typhimurium chi4550. The hSMP-1 component was localized on the surface of the head of mature rat spermatozoa by an immunofluorescence technique using polyclonal anti-hSMP-1 antibodies. Since rat sperm contain hSMP-1, this rodent can be used to assay the immunogenicity of pYA3149R. Female Wistar rats were immunized by oral administration of the recombinant Salmonella. Anti-hSMP-1 antibodies in blood and vaginal washes of immunized animals were determined. Both body fluids contained significant amounts of the antibodies, showing that the recombinant Salmonella is an effective oral immunogen in rats.
HSD-I基因编码一种人类精子膜蛋白(hSMP-1),其登录号为U12978。该基因位于人类9号染色体p12 - p13区域。当HSD-I的1.7 kb cDNA依次用EcoRI、BamHI和HindIII酶切时,形成了一个550 bp的cDNA片段,该片段编码细胞外结构域。这个片段被克隆到asd+载体pYA3149中构建pYA3149R。重组质粒用于转化鼠伤寒沙门氏菌chi4550的无毒deltacva、deltacrp、deltaasd疫苗株。通过使用多克隆抗hSMP-1抗体的免疫荧光技术,hSMP-1成分定位于成熟大鼠精子头部表面。由于大鼠精子含有hSMP-1,这种啮齿动物可用于检测pYA3149R的免疫原性。通过口服重组沙门氏菌对雌性Wistar大鼠进行免疫。测定免疫动物血液和阴道冲洗液中的抗hSMP-1抗体。两种体液中都含有大量抗体,表明重组沙门氏菌在大鼠中是一种有效的口服免疫原。