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首页> 外文期刊>Journal of Virology >Transcriptional Regulation of T4 Bacteriophage-Specific Enzymes Synthesized In Vitro
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Transcriptional Regulation of T4 Bacteriophage-Specific Enzymes Synthesized In Vitro

机译：在体外合成的T4噬菌体特异性酶的转录调节

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In contrast to dihydrofolate reductase and four other phage-specific enzymes, the initiation of deoxynucleotide kinase is essentially prevented if rifampin is added to a culture of Escherichia coli B cells within 1.5 min after infection with T4. Deoxynucleotide kinase thus belongs to a group of so-called delayed-early enzymes that is not initiated from an immediate-early promoter site. We prepared crude extracts from infected cells in a manner designed to maintain the integrity of the complexes of native, endogenous T4 DNA with bacterial structural and enzymatic units concerned with RNA synthesis. The initiation of the synthesis of the mRNA for dihydrofolate reductase, an example of an immediate-early enzyme, and deoxynucleotide kinase, a special type of delayed-early enzyme, was studied with these extracts prepared from cells infected in the absence or presence of chloramphenicol. Initiation of transcription of the dihydrofolate reductase gene is immediate when programmed by extracts made either from cells treated with chloramphenicol prior to infection (CM extracts) or from cells 3 min into the normal infection cycle (3-min extracts). However, initiation of transcription of the deoxynucleotide kinase gene programmed by CM extracts is delayed 2 min relative to the immediate initiation of transcription of the deoxynucleotide kinase gene programmed by 3-min extracts. These experiments duplicated in vitro effects of the antibiotics on the synthesis of phage-specific mRNA previously noted only in vivo.
机译：与二羟氢酸还原酶和四种其他噬菌体特异性酶相比，如果在用T4感染后1.5分钟内加入Rifampin在1.5分钟内加入Rifampin，则基本上防止了脱氧核苷酸激酶的引发。因此，脱氧核苷酸激酶属于不从立即早期启动子位点开始的一组所谓的延迟早期酶。我们以旨在维持天然内源性T4 DNA复合物与细菌结构和酶与RNA合成的酶单位的完整性的方式制备了来自感染细胞的粗提取物。对二羟氢酯还原酶的合成的合成，即直接早期酶的实例，一种特殊类型的延迟早期酶的实例，通过感染在不存在或存在的氯霉素的情况下的细胞制备的这些提取物研究。当通过在感染（CM萃取物）或3分钟内用氯霉素处理的细胞或3分钟进入正常的感染循环（3分钟提取物）时，当通过用氯霉素处理的细胞进行编程时，转录二氢氢溶胶还原酶基因的转录是立即的。然而，通过CM提取物编程的脱氧核苷酸激酶基因的转录相对于通过3分钟提取物编程的脱氧核苷酸激酶基因的直接启动延迟2分钟。这些实验对抗生素的体外效应对先前仅在体内注意的噬菌体特异性mRNA的合成中的体外影响。

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