Su Y C, Chen S L
Department of Agricultural Chemistry, National Taiwan University, Taipei, Republic of China.
Proc Natl Sci Counc Repub China B. 1996 Jul;20(3):87-91.
A genomic bank from Brevibacterium divaricatum has been prepared using lambda EMBL3 as a vector. The genomic bank's titers are 2.2 x 10(6) pfu/micrograms. Through screening by plaque hybridization, a 9.6 kb NcoI fragment which contains the entire trp operon has been isolated. Polymerase chain reaction amplification and restriction endonuclease analysis of PCR fragments indicated that there is homology between the coryneform bacteria; however, some genetic diversity among the species still exists. By complementation tests using subcloning of the 9.6 kb NcoI fragments and various E. coli tryptophan auxotrophs, this fragment was found to contain a gene cluster composed of trpE, trpD, trpC, trpB and trpA in this order. This revealed that the tryptophan biosynthesis genes in B. divaricatum may be an operon.
已使用λEMBL3作为载体构建了来自分叉短杆菌的基因组文库。该基因组文库的滴度为2.2×10⁶ 噬菌斑形成单位/微克。通过噬菌斑杂交筛选，分离出了一个包含整个色氨酸操纵子的9.6 kb NcoI片段。PCR片段的聚合酶链反应扩增和限制性内切酶分析表明，棒状杆菌之间存在同源性；然而，这些物种之间仍存在一些遗传多样性。通过使用9.6 kb NcoI片段的亚克隆和各种大肠杆菌色氨酸营养缺陷型进行互补试验，发现该片段包含一个由trpE、trpD、trpC、trpB和trpA依次组成的基因簇。这表明分叉短杆菌中的色氨酸生物合成基因可能是一个操纵子。
