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.