Study of The Transgenic Efficiency in Different Spermiogenesis Stages in Mice

In the past 30 years, the research and application of transgenic technology in gene expression of animal had become a noticeable advancement in experimental biology and applied biology. Microinjection, retrovirus mediated method and embryonic stem cells method were all the traditional methods of producing transgenic animals. But these methods had defects and limited application in the research of transgenic animals in the future. Exogenous gene was transferred into germ cells of male mice, in order to study efficiency of transgenic mice in different spermiogenesis stages. The green fluorescence protein expression plasmid(pIRES2-EGFP) and liposomes were mixed and injected into male mouse testis and epididymis. Then the intratesticular mice were mated with female at day 7, 16, 30 and 42 after infection. Polymerase chain reaction(PCR) and Southern blot were applied to identify transgenic mice. The positive ratio was 6.82%, 0, 56.86%, 42.86% by PCR analysis and 6.82%, 0, 47.06%, 34.69% by Southern blot analysis. The transgenic mice showed green fluorescence in fluorescence imager and fluorescent microscope under EGFP excitation light (488 nm). Through comparing efficiency of transgenic mice in different spermiogenesis stages, it could provided an important theoretic foundation for efficient production of transgenic animals by testis mediated gene transfer.