[Studies of foreign gene integeation during embryo early development].
There are still some problems in transgenic animals. Gene transfer, for example, reamins a difficult and costly task for animals, the vectors carrying the gene coding for the proteins of interest are of unpredictable efficiency. Therefore, it is important to identify foreign gene integration in genomose before transferring fertilized eggs to receptors, in order to increase efficiency of producing transgenic animal. In this paper, the construct that mice whey acid protein (WAP) gene promoter directs G-CSF gene was used to microinject fertilized eggs of mice. Fertilized eggs containing foreign gene were measured by using PCR method. The results showed that 100%, 77.7% and 44.4% retentions of foreign gene were achieved in 1, 2 and 8 cell-stage, respectively. Two part homologous recombination fragments were constructed and coinjected in to fertilized eggs of mice. PCR amplification fragment went beyond this homologous recombination area. If foreign gene could not integrate in to genomose, the fragment of PCR amplification could not be produced during embryo development. The results showed that the rationes of foreign gene integrated in to genomoes in 1, 2 and 8 cell-stage were 11.1%, 55.5% and 44.4%, respectively. This method might provide us a way to screen transgenic eggs when we use embryo section technique in farm animal.
Lu
Y F
YF
Institute of Biotechnology, Academy of Military Medical Science, Beijing.
Tian
C
C
Deng
J X
JX
Xiao
C Z
CZ
MA
Q J
QJ
D000818:Animals; D004622:Embryo, Mammalian / Q000378:metabolism*; D005260:Female; D008297:Male; D051379:Mice; D016133:Polymerase Chain Reaction; D011247:Pregnancy; D011995:Recombination, Genetic; D019076:Transgenes