精子介导生产转hCD59基因猪

为研究精子介导法生产异种器官移植用转基因猪的效率,将人膜反应性溶解抑制物(hCD59)基因与脂质体混合制成基因/脂质体复合物,分别采用睾丸注射法和基因脂质体复合物/精子共孵育法制备转人hCD59基因猪,其中睾丸注射法处理公猪1头,44 d后配母猪8头,7头受孕(受孕率87.5%),产仔73头,PCR阳性猪3头(阳性率4.3%);外源基因/精子共孵育法处理公猪4头,处理母猪4头,1头受孕(受孕率25%),产仔11头,PCR阳性猪1头(阳性率9%)。从15头受孕母猪共获得84只仔猪,经PCR初步检测,其中4只仔猪为转基因阳性,阳性率4.8%。研究结果表明:通过精子介导法可以获得转基因猪。

Sperm-mediated Gene Transfer to Produce hCD59 Transgenic Porcine

To study the efficiency of sperm-mediated gene transfer to produce transgenic porcine,the hCD59 gene were mixed with liposome,and two methods were used to produce transgenic porcine.Firstly,the gene/ liposome complex were directly injected into male swine's testis.The transfected male copulated with eight females 44 days later.And seven were pregnant(the pregnancy rate was 87.5%),and seventy-three piglets were got,three of them were PCR positive(the positive rate was(4.1%)).Secondly,the gene/ liposomecomplex were incubatedwith sperm,then artificially inseminated.One was pregnant(the pregnancyrate was 25%) and nine piglets were got,one of them was PCR positive(the positiverate was 9%).PCR result system indicated that the ability to produce transgenic porcine varied among different methods.According to PCR detection,the single transgenic positive rate of swine offspings was 4.8%.Double transgenic positivepiglet were not got.This result demonstrated the feasibility of sperm-mediated gene transfer to produce transgenic porcine.Compared with traditional microinjection-based gene transfer,this method is simple and valuable for other researchers to prepare transgenic animals.