RecA蛋白介导的转基因小鼠制备  

Recombinase RecA-mediated Mouse Transgenesis by Microinjection

作　　者：赵永贞[1] 肖邦[1] 张兴举[1] 唐中林[1] 崔文涛[1] 杨述林[1] 李奎[1] 

机构地区：[1]中国农业科学院北京畜牧兽医研究所

出　　处：《畜牧兽医学报》2008年第3期309-313,共5页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家“863”项目(20060110Z1039);中国农业科学院一级杰出人才科研启动基金

摘　　要：受精卵原核内核酸酶对外源基因的降解是导致显微注射法制备转基因动物效率低的重要原因之一,RecA蛋白可与单链DNA结合而阻止细胞核内核酸酶对单链DNA的降解。本试验建立了显微注射和RecA蛋白结合制备转基因动物的技术模型,并初步探讨了RecA蛋白对显微注射法制备转基因动物效率的影响。以单链DNA、RecA蛋白与单链DNA的复合体以及双链DNA为外源基因进行受精卵原核显微注射转基因试验,分别获得F0代小鼠28、41和32只,并通过PCR和Southern blot进行了转基因鉴定。结果显示,在小鼠受精卵原核显微注射Re-cA+ssDNA复合体获得的F0代中14.6%(6/41)为转基因个体,显微注射dsDNA获得的F0代中转基因个体占6.2%(2/32),而显微注射ssDNA获得的F0代中无一为转基因个体。One main reason of low efficiency producing transgenic animal by microinjection is di gestion of foreign DNA by nucleases in nuclear. Recombinase RecA can protect DNA from diges tion by phosphodiesterases or nucleases. A recombinase RecA-mediated mouse transgenesis method was developed. Further, we discussed the effect of recombinase RecA on production efficiency of transgenic mouse by microinjection. 41 F0 mice were obtained by microinjecting complexes of single strand DNA and RecA protein, Using single strand DNA and double strand DNA for microinjection, 28 and 32 F0 mice were obtained, respectively. All F0 mice were determined to be transgenic by PCR and Southern blot. The result showed that 14.6%（6/41） of F0 mice produced by pronuclear microinjection of RecA coated ssDNA was transgenic. The production efficiency of transgenic mouse was 6.20% when dsDNA was used. There was no transgenic when ssDNA was used.

关 键 词：RecA蛋白 显微注射 SSDNA 转基因 

分 类 号：Q789[生物学—分子生物学]