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我们从人睾丸组织中提取ＲＮＡ，并进一步纯化出ｍＲＮＡ；以此为模板，在反转录酶和ＤＮＡ多聚酶的作用下，合成ｃＤＮＡ；将ｃＤＮＡ与λｇＩＩ载体重组后转染大肠杆菌，构建人睾丸ｃＤＮＡ表达文库。运用遗传显色法和噬菌斑原位杂交法鉴定表达文库后，用兔抗人精子抗体筛选人精子抗原基因表达克隆（ＨＳＧ）。对ＨＳＧ表达抗原（ＨＳＧＡｇ）和特异抗体（ＨＳＧＡｂ）进行纯化和抗生育效应的测定。结果显示：（１）人睾丸ｃＤＮＡ表达文库容量为１．８２Ｘｌ０６ｐｆｕ，遗传显色法示重组率为６７％，噬菌斑原位杂交法示重组率为５１％。（２）经兔抗人精子抗体筛选２Ｘｌ０４ｐｆｕ，得８株人精子抗原基因表达克隆。（３）人血清、兔血清ＨＳＧ２Ａｂ和兔血清ＨＳＧ８Ａｂ对人精子具有补体依赖细胞毒作用。（４）兔血清ＨＳＧ３Ａｂ能阻断人精子在顶体反应时顶体后区ＣｏｎＡ受体的暴露。结果提示，ＨＳＧ２、ＨＳＧ３和ＨＳＧ８克隆的表达抗原是精子有效抗原，能作为精子免疫避孕疫苗的候选成分。
We extracted RNA from human testicular tissue and further purified the mRNA. Using this as a template, cDNA was synthesized under the action of reverse transcriptase and DNA polymerase. The cDNA was recombined with λgII vector and transfected into E. coli to construct human testis cDNA expression library. After identifying the expression library by genetic chromogenic assay and plaque in situ hybridization, human sperm antigen gene expression clone (HSG) was screened with rabbit anti-human sperm antibody. Purification and anti-fertility effects of HSG-expressing antigen (HSGAg) and specific antibody (HSGAb) were assayed. The results showed that: (1) The human testis cDNA library capacity of 1.82Xl06pfu, genetic chromosomal method showed that the recombination rate was 67%, plaque in situ hybridization showed that the recombination rate was 51%. (2) 2Xl04pfu was screened by rabbit anti-human sperm antibody to obtain 8 human sperm antigen gene expression clones. (3) Human serum, rabbit serum HSG2Ab and rabbit serum HSG8Ab have complement-dependent cytotoxic effects on human sperm. (4) Rabbit serum HSG3Ab can block the exposure of human sperm in the acrosome reaction of acrosome reaction. The results suggest that the expressed antigens of HSG2, HSG3 and HSG8 clone are sperm effective antigens, which can be used as candidate components of sperm immunization contraceptive vaccine.