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粘质沙雷氏菌编码单链DNA结合蛋白(SSB)的基因及其启动子区域的克隆与测序。

Cloning and sequencing of the Serratia marcescens gene encoding a single-stranded DNA-binding protein (SSB) and its promoter region.

作者信息

de Vries J, Wackernagel W

机构信息

Fachbereich Biologie, Universität Oldenburg, Germany.

出版信息

Gene. 1993 May 15;127(1):39-45. doi: 10.1016/0378-1119(93)90614-9.

Abstract

The gene (ssb) coding for a single-stranded DNA-binding protein (SSB) was identified on a 1.2-kb EcoRI-SalI fragment cloned from chromosomal DNA of Serratia marcescens. The cloned fragment conferred increased resistance against UV and mitomycin C (MC) to ssb- mutants of Escherichia coli. The nucleotide (nt) sequence revealed that SSB consists of 175 amino acids (aa) and has an M(r) of 18,677. It shows 89% aa sequence homology with the SSB of E. coli. The nt sequence preceding the gene contains three promoters. Two of them overlap with a presumptive SOS box, and the distal one overlaps with a second SOS box that coincides with the promoter of the adjacent uvrA (gene encoding the UvrA protein). The uvrA is transcribed in a direction opposite to that of ssb. The sequence coding for the N terminus of the UvrA of S. marcescens indicates that the first 74 aa are identical to those of the E. coli protein. The results suggest that the two bacterial SSBs are members of a group which differs from the known SSBs of prokaryotic transmissible plasmids, because their aa sequence homology with these proteins is only about 60%.

摘要

从粘质沙雷氏菌染色体DNA克隆的一个1.2kb的EcoRI - SalI片段上鉴定出了编码单链DNA结合蛋白(SSB)的基因(ssb)。该克隆片段赋予大肠杆菌的ssb - 突变体对紫外线和丝裂霉素C(MC)更强的抗性。核苷酸(nt)序列显示,SSB由175个氨基酸(aa)组成,分子量为18,677。它与大肠杆菌的SSB有89%的氨基酸序列同源性。该基因之前的核苷酸序列包含三个启动子。其中两个与一个假定的SOS框重叠,较远的一个与第二个SOS框重叠,该SOS框与相邻的uvrA(编码UvrA蛋白的基因)的启动子重合。uvrA的转录方向与ssb相反。粘质沙雷氏菌UvrA N端的编码序列表明,前74个氨基酸与大肠杆菌蛋白的相同。结果表明,这两种细菌的SSB是一个不同于原核可转移质粒已知SSB的群体成员,因为它们与这些蛋白的氨基酸序列同源性仅约为60%。

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