采用高分子介导精子作载体制备转基因泥鳅

Transgenic Loach Produced by Using Sperm Cells Mediated by High Molecules

为探讨树形高分子介导精子载体技术产生转基因动物 ,将泥鳅精子与具有标记基因LacZ的pCH1 1 0重组质粒和树形高分子在保存液内孵育 ,经DNA原位杂交检测发现树形高分子介导下精子携带外源DNA的效率得到较大的提高。将捕获了外源DNA的精子 ,再与泥鳅卵进行体外人工受精。由此发育的鱼苗经PCR和LacZ组织化学检测 ,获得了高比例的转基因泥鳅 。

The pCH110 plasmid DNA which has a marker gene LacZ controlled by a SV40 promoter was mixed with a kind of high molecule of dendrimers(cyclic core dendrimer)and then formed a DNA/dendrimer complex. Loach sperms were incubated with the DNA/dendrimer complex. Then in situ hybridization was used to detect whether the sperm cells captured the foreign DNA or not. Results show that the number of sperm capturing foreign DNA via dendrimers as vectors was obviously increased. The sperms were mixed with eggs for in vitro fertilization. PCR was used to detect the transgenic loach fries. Obvious expression of LacZ gene in the head of transgenic loach were observed by analysis through LacZ histochemical staining of embryos or fries.