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| Construction of Sleeping Beauty(SB) Transposon Vector and Its Expression in Porcine (Sus scrofa) Fetal Fibroblasts Cells |
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Abstract The sleeping beauty (SB) transposon is a Tc1/mariner family transposon. In order to examine the transposition efficiency of SB when it is used with different transposases and the same transposase at different ratios, our study constructed recombinant vector which was transfected into porcine(Sus scrofa) fetal fibroblasts cells (PEFs) and its transfection efficiency was detected by fluorescence microscope observation and qRT-PRC in an attempt to improve the transfection condition. The results of PT2-SV40-EGFP with different transposase at the same ratio of 10∶1showed that the transfection efficiency of EGFP(enhanced green fluorescent protein) was close, but the difference between expression of EGFP was significant, and SB100X has the highest expression. The results of PT2-SV40-EGFP with SB100X at different ratios showed that expression difference was significant at similar transfection efficiency. The expression level of PT2-SV40-EGFP and SB100X mixed at the ratio of 1∶1 was 50 times as high as the the group at the ratio of 5∶1 and the group at the ratio of 10∶1. This will provide technological platform for producing transgenic animals to further study SB transposon.
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Received: 06 January 2013
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