Nuclear transplantation of somatic cells of transgenic red carp (Cyprinus carpio haematopterus).
Nuclear transplantation was performed using kidney cells, tail fin cells and cultured tail fin cells of 18th passage of the F4 hGH gene transferred red carp (Cyprinus carpio haematopterus) as donors and the eggs of loach (Misgurnus anguillicaudatus) of Huanghe carp (Cyprinus carpio haematopterus) as recipients. Using loach unfertilized eggs as recipients 0.33% of reconstructed embryos developed in to neural plate from kidney cells, and 0.1% of reconstructed embryos developed in to muscular reaction stage from tail fin cells of F4hGH gene transferred red carp. A tatal of 0.4% reconstructed embryos from cultured tail fin cells of 18th passage and unfertilized eggs of Huanghe carp developed in to myotome stage. All the nuclear transfer embryos were positive for hGH gene by PCR analysis. The results suggest that exogenous hGH gene is stable in the F4hGH gene transferred red carp, hGH gene exists in all the cells of the F4 transgenic red carp. Somatic cell nuclear transfer is an effective pathway for producing homogenous transgenic fish, but with low efficiency. The reasons of low efficiency of somatic nuclear transplantation may be incompatibility of nuclear and ooplasm, incongruity of cell cycle, aberrance of chromosomes, etc.
Zhao
Hao-Bin
HB
The National Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, China. haobinzh@yahoo.com
Zhu
Zuo-Yan
ZY
D000818:Animals; D030801:Animals, Genetically Modified; D002347:Carps / Q000196:embryology* / Q000235:genetics; D002478:Cells, Cultured; D004625:Embryo, Nonmammalian / Q000378:metabolism; D047108:Embryonic Development; D005260:Female; D015870:Gene Expression; D019382:Human Growth Hormone / Q000235:genetics; D006801:Humans; D053652:Nuclear Transfer Techniques; D010063:Ovum / Q000254:growth & development* / Q000378:metabolism; D016133:Polymerase Chain Reaction
D019382:Human Growth Hormone