Generation of Disease-resistant Transgenic Chicken by Testis-mediated Injection Method of Combined Mx-NA Gene

JIN Kai;WANG Yilin;ZUO Qisheng;LI Dong;ZHANG Lei;FU Dezhi;WANG Yingjie;Zhang Yani;LI Bichun;College of Animal Science and Technology,Yangzhou University;

testis injection method;;Mx-NA combined genes;;transgenic chicken

In this study,the recombinant vectors pVITRO2-Mx-NA were injected into the testis through spermatogonialstem cell-mediated method to produce the antiviral transgenic chicken. The linearization recombinant vectors pVITRO2-Mx-NA entrapped by liposome were injected into the testis randomly,and the semen of the rooster was collected atdifferent periods(25 d,50 d,60 d,70 d),then the normal hens were artificially inseminated with the above semen,PCRtechnology and Western blot was conducted to detect the integration of the exogenous NA-Mx gene in the offspring′sblood and tissue. The results were as follows:PCR detection confirmed that exogenous Mx-NA gene had been expressedin the sperm genome,the exogenous Mx-NA gene were confirmed to be expressed in the sperm genome,the positive ratioof which was 33.33%(2/6) and the positive ratio of F1 offspring was 31.43%(11/35). Western blot results showedthat the positive rate of F1 generation was 28.6%(10/35). All above was proved that the exogenous NA-Mx genescould be integrated into the chicken genome,indicating the testis injection method would be a feasible,simple approach toproduce disease-resistant transgenic chicken.