睾丸注射Mx基因生产抗病转基因鸡初探

Transfection of Mx gene by testis-mediated method to produce disease resistance transgenic chickens

采用脂质体转染法,将线性化pcDNA3.0-MMx质粒随机打点注射入18周龄青年公鸡睾丸内,术后观察公鸡日常活动、常规检测精子;采集转染后25、50、75 d公鸡精液,并于转染70 d后人工采精,授精于正常母鸡,产生F1后代,采用常规PCR检测技术,检测证实外源Mx基因的整合情况,探讨外源Mx基因在鸡体内稳定整合和遗传的能力。结果表明:手术法转染外源基因过程对公鸡的生殖功能未造成明显影响;PCR检测证实外源Mx基因已整合到精子基因组,且能通过体外受精传递给后代,后代PCR检测的阳性率为27.27%(6/22)。证明外源Mx基因能稳定整合到基因组上,睾丸内注射法可能是一种切实可行、易于推广的制备抗病转基因鸡的方法。

The study aimed to explore the integration and hereditary ability of exogenous Mx gene in chicken by testis injection and to provide a reference for transgenic chicken production.The linearized plasmid pcDNA3.0-MMx was injected into eighteen-week-old rooster testis by lipofection method.Then daily activities of cocks were observed and semen was collected at different periods （25,50,75 d）.After 70 days from vector injection,semen was detected and used for artificial insemination into each wild-type hen.The integration of Mx gene was tested in the level of DNA by PCR.The results showed that transfected cocks displayed the normal reproductive function,and Mx gene was inserted into chicken semen genomic DNA which also can transmitted to the offspring F1.And the positive rate of PCR in the offspring F1 was 27.27%（6/22）.The data above suggest that testis-mediated is a feasible method for the preparation of transgenic chicken.