Takagi M, Uchino S, Sugimoto M, Kawai S, Hikiji T, Yano K
Department of Agricultural Chemistry, University of Tokyo, Japan.
J Basic Microbiol. 1988;28(5):335-42. doi: 10.1002/jobm.3620280508.
For the purpose of isolation of promoter regions which are regulated by a carbon source in the medium in an n-alkane-assimilating yeast, Candida maltosa, two promoter-probe vectors were constructed. Each of them consists of the LEU2 gene of Saccharomyces cerevisiae whose 5'-noncoding region was trimmed with BAL31, an autonomously replicating sequence isolated from C. maltosa genome (the TRA region) which we have previously isolated, and the pBR322 sequence. One of them, pPLC2, having the TATA box, lacks the regulatory sequence ("sequence L") of the LEU2 gene, and the other, pPLC1, lacks both the TATA box and sequence L. Using pPLC1 as a short-gun cloning vector in C. maltosa, many promoter regions which were active when glucose was present in the medium as a carbon source were obtained from the genome of C. maltosa. The sizes of the inserted fragments of two of them were determined. (In this paper, a promoter region refers to a promoter which includes a TATA box, plus a regulatory sequence such as an UAS (upstream activating sequence)-like sequence).
为了在正烷烃同化酵母麦芽糖假丝酵母中分离受培养基中碳源调控的启动子区域，构建了两种启动子探针载体。它们各自由酿酒酵母的LEU2基因组成，该基因的5'-非编码区用BAL31进行了修剪，一个从麦芽糖假丝酵母基因组中分离的自主复制序列（TRA区域）（我们之前已分离），以及pBR322序列。其中一个载体pPLC2含有TATA盒，但缺少LEU2基因的调控序列（“序列L”），另一个载体pPLC1则既缺少TATA盒也缺少序列L。使用pPLC1作为麦芽糖假丝酵母的鸟枪法克隆载体，从麦芽糖假丝酵母基因组中获得了许多在培养基中以葡萄糖作为碳源时具有活性的启动子区域。测定了其中两个插入片段的大小。（在本文中，启动子区域指的是包含TATA盒以及诸如类上游激活序列（UAS）等调控序列的启动子）。
