首页> 外文期刊>Journal of Molecular Biology >KINETICS OF EXPRESSION OF INDUCIBLE BETA-GALACTOSIDASE IN MURINE FIBROBLASTS - HIGH INITIAL RATE COMPARED TO STEADY-STATE EXPRESSION
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KINETICS OF EXPRESSION OF INDUCIBLE BETA-GALACTOSIDASE IN MURINE FIBROBLASTS - HIGH INITIAL RATE COMPARED TO STEADY-STATE EXPRESSION

机译:鼠成纤维细胞中可诱导的β-半乳糖苷酶表达的动力学-与稳态表达相比,高起始速率

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We have constructed a murine fibroblast cell line in which synthesis of beta-galactosidase can be induced by incubation with isopropyl-beta-D-thiogalactopyranoside (IPTG). This was obtained by transfection by both a plasmid expressing lacI and a second plasmid expressing lacZ from a modified simian virus 40 (SV40) promoter containing a Iac operator. We have measured the induction kinetics as well as the basal and induced differential rate of synthesis of beta-galactosidase. The steady-state rate of synthesis is tenfold higher in the presence than in the absence of inducer; we calculate an average of 1200 lacZ polypeptides are synthesized per minute per cell in the induced cultures. However, immediately after induction, the rate of accumulation of beta-galactosidase is up to 50-fold higher than the basal level. Based on our measurements of stability of beta-galactosidase, we suggest that induction may result in a subsequent down-modulation of the transcriptional activity from the induced gene. We hypothesize this inhibition may result from structural changes in DNA components, such as nucleosomes. (C) 1995 Academic Press Limited. [References: 16]
机译:我们已经构建了鼠成纤维细胞系,其中可以通过与异丙基-β-D-硫代半乳糖吡喃糖苷(IPTG)孵育来诱导β-半乳糖苷酶的合成。这是通过表达lacI的质粒和表达lacZ的第二个质粒从含有Iac操纵子的修饰的猿猴病毒40(SV40)启动子转染而获得的。我们已经测量了β-半乳糖苷酶合成的诱导动力学以及基础和诱导的差异速率。存在条件下的稳态合成速率比不存在诱导剂时的稳态速率高十倍。我们计算出诱导培养物中每细胞每分钟平均合成1200个lacZ多肽。然而,诱导后立即,β-半乳糖苷酶的积累速率比基础水平高出50倍。基于我们对β-半乳糖苷酶稳定性的测量,我们建议诱导可能导致诱导基因的转录活性发生下调。我们假设这种抑制可能是由于DNA成分(如核小体)的结构变化引起的。 (C)1995 Academic Press Limited。 [参考:16]

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