YAN Hai-fen, YI Kang-le, TREFIL P, HU Xong-gui, DENG Yuang, ZHU Li-jun
China Biotechnology. 2012, 32(12): 73-79.
There were two kinds of pluripotent stem cells in the fertile chicken eggs, one was blastodermal cells (BCs) and the other was primordial germ cells (PGCs) and both of them were the main methods of transgenic chicken. The White Leghorn (WL) and Isa Brown (IB) fertile eggs at the stage of BCs and PGCs, were windowed equatorially, and the Lentivrial vector pLenti6/v5-DEST-EGFP was microinjected into the eggs. For BCs fertile eggs from WL, the positive embryos was 64.7%(11/17)when the 13-day embryos were checked by PCR. For the PGCs (eggs incubated for72h) stage fertile eggs from IB, the hatchability was 35%, three chickens died little time after hatching, which were all positive when checked using the liver DNA by PCR at the age of 1 month and 6 month. There were three chickens among four alive ones (75%) was positive when checked the blood DNA by PCR, however they were very weak positive at the age of 12 month. For the PGCs (eggs incubated for 72~79h) stage fertile eggs from WL, the average hatchability was 21.1%, the ratio of being able to injected eggs through the window was 75.0%~92.9%,which was highest and covered the incubated time from 73h to 77h. The ratio was 44.4% when the hatched chickens were checked using the blood DNA by PCR. Comparing BCs method, the PGCs method was much better, such as on the hatchability, embryo locating on the equatorial window and so on.
