一种携带来自产博来霉素的轮状链霉菌的博来霉素N-乙酰转移酶基因作为选择标记的哺乳动物载体。

A mammalian vector carrying the bleomycin N-acetyltransferase gene from bleomycin-producing Streptomyces verticillus as a selective marker.

作者信息

Sugiyama M, Tanaka M, Kumagai T

机构信息

Institute of Pharmaceutical Sciences, Faculty of Medicine, Hiroshima University, Japan.

出版信息

J Antibiot (Tokyo). 1999 Oct;52(10):895-9. doi: 10.7164/antibiotics.52.895.

DOI:10.7164/antibiotics.52.895

PMID:10604759

Abstract

A gene, blmB, encodes a bleomycin (Bm) N-acetyltransferase, designated BAT, from Bm-producing Streptomyces verticillus and confers resistance to Bm in Streptomyces and Escherichia coli. COS-1 cells transfected with a plasmid designated pEF-BOS/blmB, in which blmB is under the control of a strong promoter from the human polypeptide chain elongation factor 1alpha, transiently produced BAT. Immuno-cytochemical analysis using an anti-BAT monoclonal antibody revealed that BAT was localized in the nucleus of the blmB-carrying COS-1 cells. NIH/3T3 cells, transfected with pEF-BOS/blmB, stably expressed BAT at least for one month. The stable transformants of blmB showed specific resistance to the Bm family of antibiotics, suggesting that blmB has potential as a selective marker in gene transfer studies with mammalian cells.

摘要

基因blmB编码一种来自产博来霉素的弗氏链霉菌的博来霉素（Bm）N - 乙酰基转移酶，命名为BAT，并赋予链霉菌和大肠杆菌对Bm的抗性。用命名为pEF - BOS/blmB的质粒转染COS - 1细胞，其中blmB受人类多肽链延伸因子1α的强启动子控制，可瞬时产生BAT。使用抗BAT单克隆抗体的免疫细胞化学分析表明，BAT定位于携带blmB的COS - 1细胞的细胞核中。用pEF - BOS/blmB转染的NIH/3T3细胞至少在一个月内稳定表达BAT。blmB的稳定转化体对博来霉素家族抗生素表现出特异性抗性，这表明blmB在哺乳动物细胞基因转移研究中具有作为选择标记的潜力。