Brurberg M B, Haandrikman A J, Leenhouts K J, Venema G, Nes I F
Laboratory of Microbial Gene Technology, Agricultural University of Norway.
Appl Microbiol Biotechnol. 1994 Oct;42(1):108-15. doi: 10.1007/BF00170232.
A chitinase gene from the Gram-negative bacterium Serratia marcescens BJL200 was cloned in Lactococcus lactis subsp. lactis MG1363 and in the silage inoculum strain Lactobacillus plantarum E19b. The chitinase gene was expressed as an active enzyme at a low level in Lactococcus lactis, when cloned in the same transcriptional orientation as the gene specifying the replication protein of the vector pIL253. Using the expression vectors pMG36e and pGKV259 with lactococcal promoter fragments p32 and p59, the expression in L. lactis was increased nine- and 27-fold, respectively. An additional twofold increase was obtained after cloning the gene under the control of p59 in the high-copy number replicon pIL253. In Lactobacillus plantarum, chitinase activity was expressed from p32, and the activity was at the same level as under p32 control in L. lactis.
将来自革兰氏阴性菌粘质沙雷氏菌BJL200的几丁质酶基因克隆到乳酸乳球菌乳酸亚种MG1363和青贮接种菌株植物乳杆菌E19b中。当几丁质酶基因以与指定载体pIL253复制蛋白的基因相同的转录方向克隆时，它在乳酸乳球菌中以低水平表达为一种活性酶。使用带有乳球菌启动子片段p32和p59的表达载体pMG36e和pGKV259，在乳酸乳球菌中的表达分别增加了9倍和27倍。在高拷贝数复制子pIL253中，将该基因克隆到p59的控制下后，表达又增加了两倍。在植物乳杆菌中，几丁质酶活性由p32表达，且该活性与在乳酸乳球菌中p32控制下的活性处于相同水平。
