[Research on spermatozoa-mediated gene transfer to product transgenic animals]

The recombinant plasmids pLNCXHLF fused with human lactoferritin gene were directly injected into male rabbit's testis and male goat's spermaductus. The transfected males were fertilized with females one month later.Using F(1)/R(1) PCR system and southern blotting,the transgenic positive rate of rabbit offsprings genomic DNA was 35%(11/31). From the PCR results of F(1)/R(1) and F(2)/R(2) system,the transgenic positive rate of genomic DNA of goat offsprings were 33.3% (4/12) and 25% (3/12) respectively. We prepared genomic DNA from 11 kinds of tissues of goat offsprings,which were tongue, lung,liver, muscle, skin,brain, gonad, spleen, intestines, heart, and kidney. The result of F(1)/R(1) PCR system indicated that the abilities to uptake exogenous DNA were various in different tissues; the positive signals of F(2)/R(2) PCR system were feebler than the ones of F(1)/R(1) PCR system,and the density of positive signals attributed to the amount of copies of exogenous DNA in the tissues. In this experiment,spermatozoa-mediated gene transfer can produce the transgenic animals after exogenous DNA being entrapped by liposome. But during the course of fertilization and the early process of embryo proliferation,the exogenous DNA had lost segments,partly integrated,or existed outside of genomic DNA. So the rate of chimera was relatively high. According to this result,this method is not only a simple and effective way to produce transgenic animals but also make references to other researchers to prepare transgenic mammals by this means.