| Fertilization is characterized by the interaction of the sperm and egg plasm membranes. Sperm membrane protein are believed to play an active role in several steps of fertilization, especially in the sperm motility, sperm-egg recognization, the fusion of gametes and the early stage of embryo development, etc. Hence, the sperm membrane proteins have gained considerable attention.In our previous studies, we have purified and characterized several kinds of membrane protein from rabbit and human sperm. Monoclonal antibodies against some sperm membrane proteins have been prepared. Using these specific monoclonal antibodies, we have isolated and identified several kinds of cDNA fragments encoding sperm membrane proteins from rat and human testis λ gt11 cDNA library by epitope selection method. Our present studies involved two main parts.1. cDNA cloning, sequencing analysis of a human sperm membrane proteinThree years ago, we obtained a cDNA fragment, named RSD-2, encoding a sperm membrane protein interacted with YWK -II monoclonal antibody from a rat testis λ gt11 cDNA expression library. In order to study the homolog in human sperm, we carried out the task of screening a human testis λ gt11 cDNA library using RSD-2 as probe. Seven positive clones hybridized with RSD-2 were isolated after the fifth cycle of screening. The size of cDNA inserts from these positive clones are:R171, 1.2kb; R321, 0.7kb; R514, 0.5kb; R642, 1.0kb;R721, 1.8kb; R915, 1.4kb; R921, 1.6kb.One of these positive clones was selected to study. This positive clone with 1.6kb insert was named R921. The insert was subcloned into pUC19 plasmid. In order to make the endonuclease map and...
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