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大肠杆菌rne基因的测序与表达

Sequencing and expression of the rne gene of Escherichia coli.

作者信息

Chauhan A K, Miczak A, Taraseviciene L, Apirion D

机构信息

Department of Molecular Microbiology, Washington University School of Medicine, St Louis, MO 63110.

出版信息

Nucleic Acids Res. 1991 Jan 11;19(1):125-9. doi: 10.1093/nar/19.1.125.

Abstract

RNase E is a major endonucleolytic RNA processing enzyme in Escherichia coli. We have sequenced a 3.2 kb EcoRI-BamHI fragment encoding the rne gene, and identified its reading frame. Upstream from the gene, there are appropriate consensus sequences for a putative promoter and a ribosome binding site. We have translated this gene using a T7 RNA polymerase/promoter system. We determined 25 amino acids from the N-terminal of the translated product and they are in full agreement with the DNA sequence. The translated product of the rne gene migrates in SDS containing polyacrylamide gels as a 110,000 Da polypeptide, but the open reading frame found in the sequenced DNA indicates a much smaller protein. The entity that migrates as a 110,000 Da contains RNA, which could account, at least partially, for the migration of the rne gene product in SDS containing polyacrylamide gels.

摘要

核糖核酸酶E是大肠杆菌中一种主要的内切核糖核酸加工酶。我们已对编码rne基因的一段3.2 kb的EcoRI - BamHI片段进行了测序,并确定了其阅读框。在该基因上游,存在适合假定启动子和核糖体结合位点的共有序列。我们使用T7 RNA聚合酶/启动子系统翻译了该基因。我们确定了翻译产物N端的25个氨基酸,它们与DNA序列完全一致。rne基因的翻译产物在含SDS的聚丙烯酰胺凝胶中以110,000 Da的多肽形式迁移,但测序DNA中发现的开放阅读框表明该蛋白要小得多。以110,000 Da形式迁移的实体含有RNA,这至少可以部分解释rne基因产物在含SDS的聚丙烯酰胺凝胶中的迁移情况。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/47d1/333542/e5d8ad48e273/nar00237-0135-a.jpg

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