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Study on Cloned Transgenic Rabbits

Author: SongWeiJie
Tutor: LiHePing；LiShanGang
School: Northeast Forestry University
Course: Animal Genetic Breeding and Reproduction
Keywords: Rabbit Somatic Cell Nuclear Transfer Transgenic Cloned Cells were transfected Vitrification GFP
CLC: S829.1
Type: Master's thesis
Year: 2011
Downloads: 59
Quote: 1
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Abstract


Donor cells and in the in vitro culture algebra have a major impact on the efficiency of somatic cell nuclear transfer. The present study by nuclear transplantation of 5-10 Low Generation and 20-25 Generation rabbit fibroblasts results found for somatic cell nuclear transfer between the two experimental groups efficiency difference exists. By donor cells after nuclear transfer reconstructed embryos fusion rate, cleavage rate, blastocyst rate and comparison of in vivo development, low-algebra is relatively efficient, high algebraic group can get a normal clone future generations. In this study verify the rabbit algebra cells in vitro cloning efficiency, provides a theoretical basis for the preparation of the gene targeting rabbit and time window. The somatic cell nuclear transfer has become a common technique to produce transgenic animals, somatic cell transfection efficiency has become an important factor of the constraints transgenic efficiency. The experiment by liposome transfection of fibroblasts cultured in vitro, was diluted at 1:40 and 1:80 density after transfection were grown to 30% -40% confluence G418 filter, and ultimately of cell clones 41, wherein the green fluorescent protein (GFP)-positive cell clones 12, two dilution density can be obtained single cell is covered with the formation of GFP clones, but the higher the efficiency of the low-density, 1:80 can be used as cell selection suitable density. The results obtained by reference to improve the success rate of cell transfection efficiency and transgenic animals. Embryo cryopreservation animal conservation has become an effective tool of the experimental animal embryo transport an essential. Vitrification is a rapid freezing method does not require special equipment. This study will screen out embryos with green fluorescent applications vitrification method for freezing and non-green fluorescent embryos screened out after vitrification. Form a complete embryo cryopreservation in vitro, 47% of the non-GFP embryonic development to the blastocyst. The recovery of GFP was gene morula ported to the receptor surrogate females body, 21% of the embryos develop into fetuses. Ultimately, four transgenic offspring pups were obtained by vitrification, and has survived for more than four months. The results showed that the GFP embryos can be successfully applied vitrification saved and fluorescent microscope filter can significantly improve the efficiency of transgenic embryos saved.

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CLC: > Agricultural Sciences > Livestock, animal medicine,hunting,silkworm,bee > Livestock > Rabbit
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