Zhao Jun, Liu Bin, Ren Wen Zhi, Zhang Shou Feng, Yu Li, Li Yuan Ping, Qiao Gui Lin, Hu Rong Liang, Yin Zhen
Military Veterinary Institute, Quartermaster University of PLA, Changchun 130062.
Shi Yan Sheng Wu Xue Bao. 2003 Jun;36(3):197-201.
The present study examined if injection of DNA into the testis tabular could generate transgenic mice via transfecting spermatogonia. The 0.9 kb Bcl-2 cDNA was fused to the promoter region of mouse whey acid protein gene and the SV40 polyA. A 3.0 kb fragment of WAP-Bcl-2-SV40 was mixed with cationic liposome and one side tabular of 3 mice testis was injected with the fragment, the other side was ligatured. Two out of the 3 males were used to mate with the female 4 days later. Twenty pups were produced and 3 of which were proven to be gene integration positive by PCR detection. Two, 1 male and 1 female, were further confirmed to carry the transgene by Southern blot analysis. The male died by accident during its feeding. The female was demonstrated to express Bcl-2 protein in its mammary glands by Western blot assay. Seven out of 45 F1 mice were proven to be transgenic by PCR. It is concluded that transfecting spermatogonia in vivo can produce transgenic mice.
本研究检测了将DNA注入睾丸曲细精管是否能通过转染精原细胞产生转基因小鼠。将0.9 kb的Bcl-2 cDNA与小鼠乳清酸蛋白基因的启动子区域及SV40 polyA融合。将3.0 kb的WAP-Bcl-2-SV40片段与阳离子脂质体混合,给3只小鼠一侧的睾丸曲细精管注射该片段,另一侧进行结扎。4天后,用其中2只雄性小鼠与雌性小鼠交配。共产下20只幼崽,经PCR检测,其中3只被证明基因整合阳性。通过Southern印迹分析进一步证实,有2只(1只雄性和1只雌性)携带转基因。雄性在饲养过程中意外死亡。通过Western印迹分析证明雌性小鼠乳腺中表达Bcl-2蛋白。经PCR检测,45只F1小鼠中有7只被证明为转基因小鼠。结论是,体内转染精原细胞可产生转基因小鼠。