【求助】Gal4的上游激活序列是什么？
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小弟最近要构建一个质粒，需要用到Gal4转录因子，有哪位大侠知道Gal4的UAS在那个质粒上？最好是容易买到的质粒啊，我查了好多质粒，都很难买到。或者能把Gal4的UAS的序列告诉我，这个上游激活序列应该不大，我可以去直接人工合成，然后参照别人那些构建成功的质粒的样子，把UAS插入到我的质粒中去，谢谢各位大侠了！！！
GAL4/UAS system
The GAL4-UAS ectopic expression system is widely used in the fruit fly, Drosophila melanogaster, for the overexpression of transgenes. This system operates under the assumption that the yeast transcription factor, GAL4, is inactive in D. melanogaster. This bipartite (two-part) expression system utilizes the yeast transcription factor GAL4 and its target sequence UAS (upstream activation sequence), to which GAL4 binds in order to activate gene transcription. GAL4 can be expressed in many different patterns by placing it under the control of various D. melanogaster promoter sequences. Since UAS promoter sequences (CGGAGTACTGTCCTCCG) are not found in D. melanogaster, it is assumed that GAL4 is inactive. This is very useful for the overexpression of transgenes from a UAS promoter because GAL4 will drive expression of the transgene while not otherwise affecting the cells.
GAL4-UAS expression systems are used to artificially express genes in organisms carrying both constructs. This is achieved by crossing a driver line with a UAS line, with the gene fused to UAS being expressed in any time and location the driver gene would normally be expressed.
References
~undefined Kramer, Jamie M., Staveley, Brian E. (2003). GAL4 causes developmental defects and apoptosis when expressed in the developing eye of Drosophila melanogaster. Genet Mol Res. 2(1):43-47. PMID 12917801
~undefined Brand, Perrimon (1993). Targeted gene expression as a means of altering cell fates and generating dominant phenotypes. Development. Jun;118(2):401-15. PMID: 8223268
The GAL4-UAS ectopic expression system is widely used in the fruit fly, Drosophila melanogaster, for the overexpression of transgenes. This system operates under the assumption that the yeast transcription factor, GAL4, is inactive in D. melanogaster. This bipartite (two-part) expression system utilizes the yeast transcription factor GAL4 and its target sequence UAS (upstream activation sequence), to which GAL4 binds in order to activate gene transcription. GAL4 can be expressed in many different patterns by placing it under the control of various D. melanogaster promoter sequences. Since UAS promoter sequences (CGGAGTACTGTCCTCCG) are not found in D. melanogaster, it is assumed that GAL4 is inactive. This is very useful for the overexpression of transgenes from a UAS promoter because GAL4 will drive expression of the transgene while not otherwise affecting the cells.
GAL4-UAS expression systems are used to artificially express genes in organisms carrying both constructs. This is achieved by crossing a driver line with a UAS line, with the gene fused to UAS being expressed in any time and location the driver gene would normally be expressed.
References
~undefined Kramer, Jamie M., Staveley, Brian E. (2003). GAL4 causes developmental defects and apoptosis when expressed in the developing eye of Drosophila melanogaster. Genet Mol Res. 2(1):43-47. PMID 12917801
~undefined Brand, Perrimon (1993). Targeted gene expression as a means of altering cell fates and generating dominant phenotypes. Development. Jun;118(2):401-15. PMID: 8223268
我查到的是这个，好像跟楼上的有点不一样？
The UAS to which Gal4 binds is CGG-N11-CCG, where N can be any base. (Campbell RN et al., 2008. Metabolic control of transcription: Paradigms and lessons from Saccharomyces cerevsiae. Biochem J 414(2):177-87)
The UAS to which Gal4 binds is CGG-N11-CCG, where N can be any base. (Campbell RN et al., 2008. Metabolic control of transcription: Paradigms and lessons from Saccharomyces cerevsiae. Biochem J 414(2):177-87)
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