Murray I A, Gil J A, Hopwood D A, Shaw W V
Department of Biochemistry, University of Leicester, U.K.
Gene. 1989 Dec 28;85(2):283-91. doi: 10.1016/0378-1119(89)90420-4.
The nucleotide sequence of a gene (cat) encoding chloramphenicol acetyltransferase (CAT) in Streptomyces acrimycini was determined. The predicted amino acid sequence demonstrates extensive homology with those of CATs isolated from Gram-negative Enterobacteria, notably with the type III variant encoded by the IncK plasmid R387. Transcript mapping indicates a single cat mRNA with a 5' end coinciding with the AUG codon used for translational initiation in vivo. We also determined the extent of a spontaneous deletion in the 5'-noncoding DNA, which occurs when the gene is cloned in the BamHI site of pBR322 in a specific orientation and which results in constitutive cat expression in Escherichia coli from the tet promoter.
测定了尖锐链霉菌中编码氯霉素乙酰转移酶(CAT)的基因(cat)的核苷酸序列。预测的氨基酸序列与从革兰氏阴性肠杆菌中分离出的CATs具有广泛的同源性,特别是与IncK质粒R387编码的III型变体。转录图谱显示有一个单一的cat mRNA,其5'端与体内翻译起始所用的AUG密码子一致。我们还确定了5'-非编码DNA中自发缺失的程度,当该基因以特定方向克隆到pBR322 的BamHI位点时会发生这种缺失,并且这会导致大肠杆菌中来自tet启动子的cat组成型表达。