腺病毒介导的高效生产水牛转基因胚胎条件的摸索

doi:10.16431/j.cnki.1671-7236.2019.03.015

Abstract

Abstract:

In order to improve the production efficiency of transgenic buffalo embryos,this study was aimed to explore the optimal conditions of adenovirus-infected buffalo granular cells and embryos.Taking buffalo granule cells and embryos developed in vitro as research objects,buffalo granular cells were infected with 0,1×10⁰,1×10^-1,1×10^-2,1×10^-3,1×10^-4,1×10^-5 GFU/mL adenovirus to obtain the best infection concentration;The buffalo granular cells were infected with the optimum concentration condition for 24,48,72 and 96 h to obtain the optimal infection time,and the results were observed by inverted fluorescence microscope.The optimal concentration and time of adenovirus-infected to buffalo granular cells were infected to the embryos in 2- and 4-cells stages,respectively,the optimal conditions of embryo infection were explored,and the embryo cleavage rate,blastocyst rate and transfection blastocyst rate were analyzed.The results showed that the optimal adenovirus infection efficiency of buffalo granular cells could be obtained by 1×10^-2 GFU/mL concentration and 48 h infection time of adenovirus.The embryos in 2- and 4-cells stages without zona pellucida and non-complete zona pellucida were infected with adenovirus got green light,while there was no light in the complete zona pellucida embryo.The embryos in 2-cell stage stopped developing after infection,while the incomplete zona pellucida embryos in 4-cell stage could continue developing into blastocyst.The blastocyst rate and blastocyst transfection efficiency of incomplete zona pellucida group was superior to other groups after infected with adenovirus in 4-cell stage.In summary, the incomplete zona pellucida,1×10^-2 GFU/mL,48 h,and 4-cell stage infection contributed to efficient expression of the target gene in buffalo embryo,and then improved the efficiency of transgenic buffalo embryo production.

Key words: adenovirus; buffalo; transgenes

CLC Number:

S814.8

LU Xingrong, PANG Chunying, DENG Tingxian, DUAN Anqin, MA Xiaoya, LIANG Shasha, LIANG Xianwei. Condition of Adenovirus-mediated High Efficient Transgenic Embryo Production in Buffalo[J]. , 2019, 46(3): 764-773.

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Condition of Adenovirus-mediated High Efficient Transgenic Embryo Production in Buffalo

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