[Introduction of foreign DNA into the genome of rare minnow by sperm electroporation].
Gene transfer was investigated in rare minnow via electroporated sperm. The sperm of the fish mixed with linear DNA (pCAhLFc) was electroporated. Then mature eggs were in vitro fertilized with these sperm cells. The DNA was extracted from the fries and analyzed by PCR. The percentage of fries with foreign genes varied between 25.5% and 66.7%. The observation of the electroporated sperm under microscope indicates that both the vigor of sperms and the successful ratio of the fertilization diminish, indicating that the electroporation with various parameters can hurt the sperm on different extent. The sperm cells electroporated with foreign genes, then was treated by DNA, and the DNA was extracted and was analyzed by PCR. It was found that foreign gene still exists, which approves that the electroporation can force the foreign gene enter the sperm of rare minnow.
Zhong
Jia-Yu
JY
Institute of Hydrobiology, Chinese Academy of Sciences, State Key Laboratory of Freshwater Ecology and Biotechnology, Wuhan 430072, China.
Mao
Wei-Feng
WF
Zhu
Zuo-Yan
ZY
D000818:Animals; D001723:Birth Rate; D003530:Cyprinidae / Q000235:genetics*; D004274:DNA, Recombinant / Q000235:genetics* / Q000494:pharmacology; D018274:Electroporation / Q000379:methods; D005307:Fertilization in Vitro / Q000379:methods; D008297:Male; D013094:Spermatozoa / Q000187:drug effects / Q000502:physiology*; D014170:Transformation, Genetic
D004274:DNA, Recombinant