Construction Of The PIRES2-EGFP-DAZ3 Plasmid And Replication And Expression Of Human DAZ Gene In Two-Cell Embryo Of Golden Hamster

[BACKGROUND & AIM] The incidence of infertility is about 10%-15%, 50% of that is caused by the male. In the male infertility, azoospermia and oligospermia frequently occurred. In 1976, Tiepolo etc. found that there is the lack of the gene which is related to the generation of sperm at the end of the Y-chromosome in six azoospermia patients, which is called azoospermia factor. Later, many scholars located this factor at the first zone of long arm of Y-chromosome through the molecule cell genetics technology. AZF is divided into three non-overlap deleted subregion AZFa, AZFb and AZFc. Recently, Kent-First etc. found that there is the fourth AZF area between the AZFb and the AZFc, which is called AZFd. Afterward, Kobayashi etc. separated the DAZ gene by, STS (the Sequence Tagged Site). Through more and more researches, the whole or partial lack of the DAZ gene cluster in the Y-chromosome is the molecular basis of the dyszoospermia. Besides the Sertoli cell only syndrome of the patients lack of DAZ, some patients can generate little sperm or spermatogone. DAZ gene is related to the generation of sperm, it is one of the important candidate gene of AZF. Moreover, 10%-15% patients of primarily azoospermia and severity oligosperm are totally lack the DAZ gene. This thesis through constructing the pIRES2-EGFP-DAZ3 plasmid, than transfected the recombinant plasmid into the NIH3T3 cell of mice, detected the expression of the recombinant plasmid in the eukaryon cell; At the same time, through transfected the sperms of the golden hamster, fertilized them with the normal oocyte after selection, and detected the replication and the expression of DAZ3 gene in the eggs, which probed the new approaches to heal the male infertility through trans-gene,...