人G-CSF转基因鼠的稳定遗传与表达

利用人粒细胞集落刺激因子（Ｇ－ＣＳＦ）基因组基因作为目的片段，将其受控于２．６ｋｂ的小鼠乳清酸蛋白（ＷＡＰ）基因的调控区下，通过显微注射法获得了两只整合有人Ｇ－ＣＳＦ转基因小鼠，通过繁殖建立了稳定的转基因系．一些表型参数测定表明转基因鼠与正常鼠无明显差别．通过ＲＴ－ＰＣＲ及Ｓｏｕｔｈｅｒｎｂｌｏｔ检测，在乳腺表达出人Ｇ－ＣＳＦ，为乳腺表达外源蛋白质及今后大动物研究奠定了基础．

Human G CSF Transgene Stable Transmission and Expression in Mice

Transgenic animals are being used for the production of valuable proteins.The strategy is to target expression to the mammary gland and harvest the product from milk.It is important to research transgene transmission and expression using mice as models.A hybrid gene that mice whey acid protein (WAP) gene 5′region directed human granulocyte colony stimulating factor (G CSF) genomic DNA was constructed to produce transgenic mice.Two transgenic mice were obtained by microinjecting 1080 fertilized eggs of mice and identified by PCR and Southern blot.Integration rate of foreign gene got 2 37%.Transgenic mice lines were established by mating with no transgenic mice and offspring were identified by PCR so as to research transgene transmission.Some parameters included body weight and some blood components such as red blood cell,thrombocyte,neutrocyte and acidophil white blood cell were determined in transgenic and normal mice.The result showed that there was no difference between transgenic and normal mice in these parameters.This suggested that these transgenic mice develop normally and transgene could transmit to next generation stable.In order to understand transgene expression,total RNA were extracted from mammary gland and transgene mRNA determined by RT PCR and Southern blot.The results showed that there was human G CSF expressed in mammary gland of transgenic mice.