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用基因重组技术构建人钙调素基因Ⅲ（ｈＣａＭⅢｃＤＮＡ）表达载体ｐＢＶ／ｈＣａＭⅢ，并在大肠杆菌ＤＨ５α中经热诱导获得可溶性ＣａＭ蛋白的高效表达．将纯化的重组ｈＣａＭ与异型双功能剂ＳＰＤＰ及鼠血清白蛋白（ＭＳＡ）交联，免疫Ｂａｌｂ／ｃ小鼠，用常规细胞融合技术制备单克隆抗体（ＭｃＡｂ），得到３株稳定分泌单克隆抗体的杂交瘤细胞．间接ＥＬＩＳＡ和斑点免疫结果证实，三种单克隆抗体均与自制的ｒｈＣａＭ和ＣａＭ标准品起特异反应．用免疫组化法对精子中ＣａＭ定位，发现ＣａＭ主要分布于精子头颈部，不育组ＣａＭ＋精子率（（４５．０±７．５）％）显著低于生育组（（６８．５±１０．５）％）．
The recombinant plasmid pBV / hCaMⅢ was constructed by gene recombination technique, and its expression was induced by heat in E. coli DH5α. The purified recombinant hCaM was cross-linked with the heterobifunctional SPDP and mouse serum albumin (MSA), Balb / c mice were immunized and monoclonal antibodies (McAbs) were prepared by conventional cell fusion techniques. Three stable monoclonal antibodies secreting Of hybridoma cells. Indirect ELISA and dot immunization demonstrated that all three monoclonal antibodies reacted specifically with homemade rhCaM and CaM standards. Using immunohistochemistry to locate CaM in sperm, we found that CaM mainly distributed in sperm head and neck. The CaM + sperm rate in sterile group (45.0 ± 7.5%) was significantly lower than that in reproductive group (68.5 ± 10.5)%).