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| The Enhanced Green Fluorescent Protein Gene(EGFP) Transfer Mediated by Sleeping Beauty Transposon in Chicken(Gallus domesticus) Embryo |
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Abstract To explore the effect of transfection of the transposon-mediated technology in the chick embryo, the single plasmid of Sleeping Beauty(SB) transposable vector pT2-SV40-EGFP-SB11 was injected into 36 h incubated chicken(Gallus domesticus) blastoderm with eggs windowing, at the same time window non-injection group and non-window group were set as control group. Stereological fluorescence microscopy and RT-PCR were used to detect and compare the expression of enhanced green fluorescent protein gene(EGFP), hatching rate, survival rate and egg weight change at different stages of embryonic. The results showed that the transposon-mediated EGFP in chicken embryos obtained a higher expression rate, in embryos of early development(6 d: 89%, 10 d: 71%), but decreased slightly during later period (14 d: 41%, 18 d: 45%, 21 d: 40%); The difference of three groups of eggs weight change on 7 d and 12 d ,was not significant(P>0.05), the egg weight ratio of the window injection group on 17 d and 21 d, was significantly lower than that of non-window group(P<0.05); the difference of survival rate among three groups on 2 d was not significant(P>0.05); window injection group on the 4 d was significantly lower than that of the window non-injection group(P<0.05), while the window non-injection group was significantly lower than that of the non-window group(P<0.05); from 6 d to 21 d, the survival rate and hatching rate among the three groups showed significant differences(P<0.01). The preliminary experiments proved that the improved egg fenestration can be used for the chick embryo transgenic research and initially establish the method of the transfection exogenous gene in chick embryo mediated with SB transposon.
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Received: 06 August 2012
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HAN Xue-Qing, GAO Feng-Ying, LU Mai-Xin, LIU Zhi-Gang, CAO Jian-Meng, WANG Miao, YI Meng-Meng, ZHANG De-Feng. Cloning, Expression and Functional Analysis of TRAF4 Gene in Nile Tilapia (Oreochromis niloticus)[J]. 农业生物技术学报, 2019, 27(3): 381-392. |
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