共查询到20条相似文献,搜索用时 15 毫秒
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Wenfang Liu Geping Wang Alexander G Yakovlev 《The Journal of biological chemistry》2002,277(10):8273-8278
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人细胞周期相关激酶启动子的克隆和初步分析 总被引:1,自引:0,他引:1
克隆人的细胞周期相关激酶(CCRK)基因启动子,分析与其表达有关的转录调控因子。通过巢式PCR方法,从人的基因组总DNA中分离出CCRK基因5′端非翻译区大小为1072bp的片段。这段片段的3′端起始点在第一个外显子里第一个翻译密码子ATG( 1)上游128bp处。以1072bp为模板,对启动子进行5′端删除分析,分别扩增951bp(-1072/-128)、564bp(-692/-128)、313bp(-441/-128)、127bp(-239/-128)的片段,与pGL3-Basic荧光素酶报告载体重组构建,人工加上克隆位点,5′端带有KpnI、3′端带有XhoI位点。瞬时转染恶性神经胶质瘤细胞株U373,通过双荧光素酶活性进行分析。对分离出的1072bp的片段进行测序,结果与GenBank(Accession AF035013)的序列比较,同源序列达到98%。双荧光素酶活性分析564bp片段有最强的活性,313bp片段为有活性的最小片段。本研究为进一步研究分析CCRK的核心启动子和与其相关的转录因子奠定基础。 相似文献
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Furukawa K Horie M Okutomi K Sugano S Furukawa K 《Biochimica et biophysica acta》2003,1627(2-3):71-78
Human GD3 synthase gene consisted of five exons and span about 135 kilobases. The 5'-flanking region lacked canonical TATA and CAAT boxes, but contained SP1 binding site(s) as in rat and mouse. The promoter activity in the 5'-flanking region (-2262 approximately +1) became definite when SV40 enhancer was added to the reporter plasmid. Luciferase assay with deletion mutants suggested the existence of a silencer region between -2262 and -978 nt similarly with those in mouse and rat. They also commonly contained a GT/CG repeat sequence at upstream of -1200 approximately -1300 nt, suggesting that they form Z-type DNA, and are involved in the gene regulation. 相似文献
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Daniela Alberati-Giani Andrea M. Cesura Clemens Broger William D. Warren Stephan Rver Pari Malherbe 《FEBS letters》1997,410(2-3)
Kynurenine 3-monooxygenase, an NADPH-dependent flavin monooxygenase, catalyses the hydroxylation of
-kynurenine to
-3-hydroxykynurenine. By hybridization screening using a cDNA probe encoding the entire exon 2 of Drosophila melanogaster kynurenine 3-monooxygenase, we isolated a 2.0 kb cDNA clone coding for the corresponding human liver enzyme. The deduced amino acid sequence of the human protein consists of 486 amino acids with a predicted molecular mass of 55 762 Da. Transfection of the human cDNA in HEK-293 cells resulted in the functional expression of the enzyme with kinetic properties similar to those found for the native human protein. RNA blot analysis of human tissues revealed the presence of a major mRNA species of 2.0 kb in liver, placenta and kidney. 相似文献
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