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由Tn1331编码的I b型氨基糖苷6'-N-乙酰基转移酶均匀分布于细胞质中。

The aminoglycoside 6'-N-acetyltransferase type Ib encoded by Tn1331 is evenly distributed within the cell's cytoplasm.

作者信息

Dery Ken J, Søballe Britta, Witherspoon Mavee S L, Bui Duyen, Koch Robert, Sherratt David J, Tolmasky Marcelo E

机构信息

Department of Biological Science, College of Natural Science and Mathematics, California State University Fullerton, Fullerton, California 92834-6850, USA.

出版信息

Antimicrob Agents Chemother. 2003 Sep;47(9):2897-902. doi: 10.1128/AAC.47.9.2897-2902.2003.

Abstract

The multiresistance transposon Tn1331, which mediates resistance to several aminoglycosides and beta-lactams, includes the aac(6')-Ib, aadA1, bla(OXA-9), and bla(TEM-1) genes. The nucleotide sequence of aac(6')-Ib includes a region identical to that of the bla(TEM-1) gene. This region encompasses the promoter and the initiation codon followed by 15 nucleotides. Since there were three possible translation initiation sites, the amino acid sequence at the N terminus of the aminoglycoside 6'-N-acetyltransferase type Ib [AAC(6')-Ib] was determined and was found to be SIQHF. This result indicated that aac(6')-Ib includes a translational fusion: the first five amino acids of the leader peptide of the TEM beta-lactamase are fused to the rest of the AAC(6')-Ib protein. This gene fusion could have formed during the genesis of Tn1331 as a consequence of the generation of a 520-nucleotide duplication (M. E. Tolmasky, Plasmid 24:218-226, 1990). An identical gene isolated from a Serratia marcescens strain has been previously described (G. Tran van Nhieu and E. Collatz, J. Bacteriol. 169:5708-5714, 1987). Extraction of the periplasmic proteins of E. coli harboring aac(6')-Ib by spheroplast formation showed that most of the AAC(6')-Ib protein is present in the cytoplasm. A genetic fusion to phoA confirmed these results. AAC(6')-Ib was shown to be evenly distributed inside the cell's cytoplasm by fluorescent microscopy with an AAC(6')-Ib-cyan fluorescent protein fusion.

摘要

介导对多种氨基糖苷类抗生素和β-内酰胺类抗生素耐药的多耐药转座子Tn1331包含aac(6')-Ib、aadA1、bla(OXA-9)和bla(TEM-1)基因。aac(6')-Ib的核苷酸序列包含一个与bla(TEM-1)基因相同的区域。该区域包括启动子和起始密码子以及随后的15个核苷酸。由于存在三个可能的翻译起始位点,因此确定了氨基糖苷6'-N-乙酰基转移酶Ib型[AAC(6')-Ib] N端的氨基酸序列,结果为SIQHF。这一结果表明aac(6')-Ib包含一个翻译融合:TEMβ-内酰胺酶前导肽的前五个氨基酸与AAC(6')-Ib蛋白的其余部分融合。这种基因融合可能是在Tn1331的发生过程中由于产生了一个520个核苷酸的重复而形成的(M. E. Tolmasky,《质粒》24:218 - 226,1990)。先前已描述了从粘质沙雷氏菌菌株中分离出的一个相同基因(G. Tran van Nhieu和E. Collatz,《细菌学杂志》169:5708 - 5714,1987)。通过原生质球形成法提取携带aac(6')-Ib的大肠杆菌周质蛋白表明,大多数AAC(6')-Ib蛋白存在于细胞质中。与phoA的基因融合证实了这些结果。通过使用AAC(6')-Ib - 青色荧光蛋白融合体的荧光显微镜观察表明,AAC(6')-Ib在细胞细胞质内均匀分布。

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