Kooistra J, Vosman B, Venema G
Department of Genetics, University of Groningen, Haren, The Netherlands.
J Bacteriol. 1988 Oct;170(10):4791-7. doi: 10.1128/jb.170.10.4791-4797.1988.
By insertional mutagenesis, 36 transformation-deficient, mitomycin C-sensitive Bacillus subtilis mutants were isolated, 16 of which were ATP-dependent DNase (ADD) deficient. PBS1 transduction showed that the mutations were closely linked to the metD marker and weakly linked to the glyB marker. With the aid of one of the mutants, a transcription unit involved in ADD synthesis was cloned. The chromosomal location of the transcription unit was established at the restriction site level by determining the presence or absence of ADD in transformants of wild-type cells obtained with various DNA fragments inserted in pUC derivatives. The transcription unit complemented a mutant in which the add transcription unit had been deleted.
通过插入诱变,分离出36个转化缺陷型、丝裂霉素C敏感的枯草芽孢杆菌突变体,其中16个是ATP依赖性DNA酶(ADD)缺陷型。PBS1转导表明,这些突变与metD标记紧密连锁,与glyB标记弱连锁。借助其中一个突变体,克隆了一个参与ADD合成的转录单元。通过确定用插入pUC衍生物的各种DNA片段获得的野生型细胞转化体中ADD的有无,在限制位点水平上确定了转录单元的染色体定位。该转录单元互补了一个缺失add转录单元的突变体。