San Francisco M J, Hope C L, Owolabi J B, Tisa L S, Rosen B P
Biotechnology Center, Ohio State University, Columbus 43210.
Nucleic Acids Res. 1990 Feb 11;18(3):619-24. doi: 10.1093/nar/18.3.619.
The regulatory region of the plasmid-encoded arsenical resistance (ars) operon was cloned as a 727-bp EcoRI-HindIII fragment. When cloned into a promoter probe vector this fragment conferred arsenite inducible tetracycline resistance in Escherichia coli, indicating that the fragment carried a regulatory gene, the arsR gene. A single region corresponding to -35 and -10 promoter recognition sites was identified. The transcriptional start site of the mRNA was determined by primer extension. The sequence has an open reading frame for a potential 13,179 Da polypeptide, termed the ArsR protein. The fragment was cloned into a temperature regulated expression vector. A protein with an apparent molecular mass of about 12 kDa was induced by either temperature or arsenite. This protein was purified and used to produce antibodies specific for the ArsR protein.
质粒编码的抗砷（ars）操纵子的调控区被克隆为一个727bp的EcoRI-HindIII片段。当将该片段克隆到启动子探针载体中时，它赋予大肠杆菌亚砷酸盐诱导的四环素抗性，这表明该片段携带一个调控基因，即arsR基因。鉴定出了一个与-35和-10启动子识别位点相对应的单一区域。通过引物延伸确定了mRNA的转录起始位点。该序列具有一个潜在的13179Da多肽的开放阅读框，称为ArsR蛋白。该片段被克隆到温度调控表达载体中。一种表观分子量约为12kDa的蛋白质可由温度或亚砷酸盐诱导产生。该蛋白质被纯化并用于制备针对ArsR蛋白的特异性抗体。
