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Abstract To compare the efficiency of transposon-mediated gene trap among SB(Sleeping Beauty), PB(PiggyBac) and Tol2, a promoter trap vector based on those transposons named pT3-PST-Pro-Trap was constructed. Then the vector plasmid DNA and transposase mRNA of SB, PB and Tol2 according to 1.0∶1.5 ratio were mixed, respectively, and injected into zebrafish(Danio rerio) one-cell embryos. The gene trapping efficiency could be determined by the expression level of green fluorescent protein report gene (GFP). The results showed that promoter trap vector mediated by SB, PB, and Tol2 transposon could capture the endogenous gene promoter with high efficiency of 24.32%、30.7% and 18.87% 36 h after injected, respectively, and GFP expression were observed. PB-mediated gene trap efficiency was significantly higher than that of SB and Tol2 (P＜0.05). Three kinds of transposon capture efficiency in 60 h period were higher than that of 36 h period, SB and Tol2 group were significant differences (P＜0.05), while PB group was no significant difference (P＞0.05). The results showed that PB, SB and Tol2 transposon mediated gene capture technology can be used to vertebrate high-throughput screening functional genes, and provide effective new method for the research of functional genes.
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Received: 27 December 2013
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YANG Han, LI Yan, SHEN Chang-Wei, JIN Yu-Meng, SHI Xiao-Qian, XIE Chang-Yan, Mei Xin-Lan, XU Yang-Chun, DONG Cai-Xia. Cloning and Expression Analysis of Potassium Channel Gene PbAKT1 in Pear (Pyrus betulifolia)[J]. 农业生物技术学报, 2019, 27(8): 1341-1350. |
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