共查询到20条相似文献,搜索用时 31 毫秒
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J D Gralla 《Proceedings of the National Academy of Sciences of the United States of America》1985,82(10):3078-3081
A DNase protection technique is described and applied to the interaction of three lac control proteins with supercoiled lac DNA. The technique uses end-labeled oligonucleotide primers to probe specific DNA regions as an alternative to protocols requiring restriction endonuclease cleavage or blotting. Thus DNA may be probed with high resolution in its native state. It is demonstrated that the introduction of supercoiling into DNA accelerates the rate of lac ps promoter binding by RNA polymerase but does not alter the positions at which polymerase, c-AMP-binding protein, or lac repressor bind to lac DNA. 相似文献
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An initiation element in the yeast CUP1 promoter is recognized by RNA polymerase II in the absence of TATA box-binding protein if the DNA is negatively supercoiled
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Leblanc BP Benham CJ Clark DJ 《Proceedings of the National Academy of Sciences of the United States of America》2000,97(20):10745-10750
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Devendra B. Srivastava Katherine Leon Joseph Osmundson Ashley L. Garner Leslie A. Weiss Lars F. Westblade Michael S. Glickman Robert Landick Seth A. Darst Christina L. Stallings Elizabeth A. Campbell 《Proceedings of the National Academy of Sciences of the United States of America》2013,110(31):12619-12624
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Construction of a plasmid that overproduces the large proteolytic fragment (Klenow fragment) of DNA polymerase I of Escherichia coli. 总被引:15,自引:9,他引:15
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C M Joyce N D Grindley 《Proceedings of the National Academy of Sciences of the United States of America》1983,80(7):1830-1834
Using currently available gene fusion techniques, we have constructed plasmids that direct the overproduction of the carboxyl-terminal two-thirds of DNA polymerase I, corresponding to the proteolytically derived "Klenow fragment." We have obtained overproduction amounting to several percent of the cellular protein using constructs in which expression is directed either from the lac promoter or from the leftward promoter of phage lambda. The polymerase fragment has been purified to homogeneity from such overproducing strains by a rapid three-stage purification procedure, yielding material capable of carrying out the same reactions (polymerization, 3' labeling, DNA sequence analysis) as the proteolytically derived fragment. The availability of such overproducing strains should greatly facilitate structural and mechanistic studies of DNA polymerase I. Moreover, the techniques we have described for the cloning and expression of a gene fragment should be generally applicable for the study of protein structure and function in other systems. 相似文献
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