Weng S F, Shieh M Y, Lai F Y, Shao Y Y, Lin J W, Tseng Y H
Institute of Molecular Biology, National Chung Hsing University, Taiwan, Republic of China.
Biochem Biophys Res Commun. 1996 Nov 12;228(2):386-90. doi: 10.1006/bbrc.1996.1671.
A broad-host-range promoter-probing vector, pMY3 (8.0 kb), was constructed for cloning of DNA fragments containing promoter sequences of Xanthomonas campestris pv. campestris. This vector (pMY3) consists of the RK2 replicon, promoterless luxAB genes, the thr attenuator to block the transcription of RNA into the luxAB region, and multiple cloning sites for cloning of the fragment carrying promoter sequences. The feasibility of using pMY3 as a promoter-probing vector in both E. coli and Xc17 was demonstrated by using the lac promoter of E. coli, and the amy promoter of X. campestris in Xc17. Among the 63 promoter-containing fragments cloned from Xc17, only 9 were able to express in E. coli. It appears that X. campestris can recognize most E. coli type promoters, but, E. coli can recognize only a small portion of the X. campestris type promoters.
构建了一种广宿主范围的启动子探测载体pMY3（8.0 kb），用于克隆含有野油菜黄单胞菌野油菜致病变种启动子序列的DNA片段。该载体（pMY3）由RK2复制子、无启动子的luxAB基因、用于阻断RNA转录进入luxAB区域的苏氨酸弱化子以及用于克隆携带启动子序列片段的多个克隆位点组成。通过在大肠杆菌中使用大肠杆菌的lac启动子以及在Xc17中使用野油菜黄单胞菌的淀粉酶启动子，证明了pMY3作为启动子探测载体在大肠杆菌和Xc17中的可行性。在从Xc17克隆的63个含启动子的片段中，只有9个能够在大肠杆菌中表达。看来野油菜黄单胞菌能够识别大多数大肠杆菌类型的启动子，但是大肠杆菌只能识别一小部分野油菜黄单胞菌类型的启动子。
