摘要
精子疫苗的研制和用ELISA法检测抗精子抗体均需用大量有效的精子抗原,传统的用物理、化学和单克隆抗体亲和层析方法直接从精子上提取有限的抗原已远远不能满足需要,我们在构建了人睾丸组织cDNA文库的基础上,用兔抗人精子血清筛选该文库,初筛了2×10~4个重组λgt_(11),发现了10个阳性噬菌斑。经挑斑扩增后复筛,其中8个仍为阳性,证明为人精子抗原的基因表达克隆。用这些基因克隆,我们可以大量制备精子抗原,从而为精子疫苗的研制和抗精子抗体ELISA检测药盒的大量制备创造了条件。
The research of sperm contraceptive vaccine and application of EL1SA for the detection of antisperm antibody need a lot of sperm antigens. Traditionally, Sperm antigens were extracted directly from the sperm by using physical and chemical methods, or using antisperm antibodies to conugate the antigens. Such obtained sperm antigens are too less to satisfy the needs. After having constructed human testis cDNA library, we screened human cloning sperm antigen genes from it by using rabbit's anti-human sperm sera as probe, we primarily screened 100 pfu of recombinant λgt11and found ten positiv plaques. Then after repeated screening only eight clones were still positive which were λgt11 recombinanted with human sperm genes. Using these sperm genes , we can get a lot of sperm antigens which can satisfy the need of sperm vaccine research and wildly application of ELISA method for antisperm antibodies.
关键词
精子
睾丸
CDNA文库
antisperm antibodies, cDNA library, human sperm antigens.
