用显微注射法建立转bcl-x_l基因小鼠  

Developing bcl-x_l transgenic mice by microinjection

作　　者：肖文伍[1] 王芙蓉 张苏明[1] 赵浩斌[2] 郑新明[2] 樊俊华[2] 魏庆信[2] 张旻 

机构地区：[1]华中科技大学同济医学院附属同济医院神经科,湖北武汉430030 [2]湖北省农业科学院生物工程室,湖北武汉430067

出　　处：《中国神经科学杂志》2002年第3期630-633,共4页

基　　金：国家自然科学基金重点项目 (3 973 0 170 ) ;国家自然科学基金面上项目 (3 9770 810 )

摘　　要：建立转bcl xl基因小鼠 ,继而传代建系 ,用于缺血性脑血管疾病的研究。采用显微注射法 ,将人bcl xl基因注入昆明小白鼠受精卵获得子代鼠 ,然后作PCR ,Southern blot ,mRNA ,Western blot检测以获得阳性鼠。实验中注射受精卵 16 5 4枚 ,移植卵数 14 4 8枚 ,受体鼠 4 9只 ,怀孕鼠 7只 ,子代鼠 13只 ,整合数 4只并均有表达 ,植入受精卵的存活率 83% ,受精卵的总存活率 0 .78% ,移植鼠的怀孕率 14 .3% ,整合效率 30 .7% ,总有效率为 0 .2 4 %。初步获得了转bclIt was attempted to develop bcl x l transgenic mouse and produce its lineages for studying ischemic cerebrovascular diseases,bcl x l gene was introduced into mice by microinjection .The presence of bcl x l and its expression were confirmed by PCR,Southern blot,mRNA inspection,Western blot in the founders.In our experiments, bcl x l gene was injected into pronucleus of 1 654 zygotes and 1 448 injected eggs were implanted into 49 foster mothers, 7 of which were pregnant and bore 13 offsprings. Among the babies, 4 offsprings were integrated with the bcl x l gene and expressed it. Totally, implanted zygotes survival rate (83%), zygotes total survival rate (0.78%), recipient mouse pregnance rate ( 14.3% ), integration rate (30.7%), expression rate(100%) ,total efficiency(0.24%)were obtained respectively. Bcl x l transgenic mice were obtained, 4 in number, but the transgenic efficiency was very low and the expression was unstable. The model of cerebral vascular disease will be made after acquiring transgenic lineages.

关 键 词：显微注射法 转bcl-x1基因 小鼠 脑缺血 动物模型 

分 类 号：R743.31[医药卫生—神经病学与精神病学]