用显微注射法建立转bcl-x_l基因小鼠

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bcl xl基因小鼠

Developing bcl-x_l transgenic mice by microinjection

It 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.