农业生物技术学报2013，Vol.21Issue(6)：684-689,6.
睡美人转座子真核表达载体的构建及在猪胎儿成纤维细胞系中的表达验证
Construction of Sleeping Beauty(SB) Transposon Vector and Its Expression in Porcine (Sus scrofa) Fetal Fibroblasts Cells
摘要
Abstract
The sleeping beauty (SB) transposon is a Tcl/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∶ 1 showed 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.关键词
睡美人(sleeping beauty,SB)/真核表达载体/转基因动物Key words
Sleeping beauty transposon/ Eukaryotic expression vector/ Transgenic animals引用本文复制引用
周家庆,王升智,宋成义,高波,李奎,钱丽丽,崔文涛,牟玉莲,李碧春..睡美人转座子真核表达载体的构建及在猪胎儿成纤维细胞系中的表达验证[J].农业生物技术学报,2013,21(6):684-689,6.基金项目
中国博士后科学基金特别资助项目(No.201104168)、国家自然科学基金(No.31200920)和国家转基因新品种培育重大专项(No.2011ZX08011-006) （No.201104168）
