0引言
蛋白质和蛋白质的相互作用是很多生命现象的基础.随着分子生物学技术的发展,特别是人类基因组计划的完成,使人类对基因的结构和功能的认识不断加深,但基因编码的蛋白质的功能研究尚是一个难题.酵母双杂交(yeast two hybrid)技术...0引言
蛋白质和蛋白质的相互作用是很多生命现象的基础.随着分子生物学技术的发展,特别是人类基因组计划的完成,使人类对基因的结构和功能的认识不断加深,但基因编码的蛋白质的功能研究尚是一个难题.酵母双杂交(yeast two hybrid)技术是利用酵母遗传学方法分析蛋白质之间的相互作用,该方法建立以来,经过不断的完善和发展,不但可以检测已知蛋白质之间的相互作用,更重要的在于发现新的与已知蛋白相互作用的未知蛋白.展开更多
The piggyBac transposon has been long used to integrate foreign DNA into insect genomes.However,undesirable transgene expression can result from random insertions into the genome.In this study,the efficiency of chimer...The piggyBac transposon has been long used to integrate foreign DNA into insect genomes.However,undesirable transgene expression can result from random insertions into the genome.In this study,the efficiency of chimeric Gal4-piggyBac transposase in directing integration onto a DNA target plasmid was evaluated in cultured silkworm Bombyx mori Bm-12 and fruit fly Drosophila Schneider 2(S2) cells.The Gal4-piggyBac transposase has a Gal4 DNA-binding domain(DBD),and the target plasmid has upstream activating sequences(UAS) to which the Gal4 DBD can bind with high affinity.The results indicate that,in the Bm-12 and S2 cells,transpositional activity of Gal4-piggyBac transposase was increased by 4.0 and 7.5 times,respectively,compared to controls,where Gal4-UAS interaction was absent.Moreover,the Gal4-piggyBac transposase had the ability of directing piggyBac element integration to certain sites of the target plasmid,although the target-directing specificity was not as high as expected.The chimeric piggyBac transposase has the potential for use in site-directed transgenesis and gene function research in B.mori.展开更多
文摘0引言
蛋白质和蛋白质的相互作用是很多生命现象的基础.随着分子生物学技术的发展,特别是人类基因组计划的完成,使人类对基因的结构和功能的认识不断加深,但基因编码的蛋白质的功能研究尚是一个难题.酵母双杂交(yeast two hybrid)技术是利用酵母遗传学方法分析蛋白质之间的相互作用,该方法建立以来,经过不断的完善和发展,不但可以检测已知蛋白质之间的相互作用,更重要的在于发现新的与已知蛋白相互作用的未知蛋白.
基金supported by the National Basic Research Program (973) of China (No. 2009CB119200)the Natural Science Foundation of Zhejiang Province (No. Y304122),China
文摘The piggyBac transposon has been long used to integrate foreign DNA into insect genomes.However,undesirable transgene expression can result from random insertions into the genome.In this study,the efficiency of chimeric Gal4-piggyBac transposase in directing integration onto a DNA target plasmid was evaluated in cultured silkworm Bombyx mori Bm-12 and fruit fly Drosophila Schneider 2(S2) cells.The Gal4-piggyBac transposase has a Gal4 DNA-binding domain(DBD),and the target plasmid has upstream activating sequences(UAS) to which the Gal4 DBD can bind with high affinity.The results indicate that,in the Bm-12 and S2 cells,transpositional activity of Gal4-piggyBac transposase was increased by 4.0 and 7.5 times,respectively,compared to controls,where Gal4-UAS interaction was absent.Moreover,the Gal4-piggyBac transposase had the ability of directing piggyBac element integration to certain sites of the target plasmid,although the target-directing specificity was not as high as expected.The chimeric piggyBac transposase has the potential for use in site-directed transgenesis and gene function research in B.mori.
