地衣芽孢杆菌sipS基因的强启动子的克隆与鉴定

The Cloning and Identification of Bacillus Licheniformis Strong Promoter

通过基因打靶的方法,利用大肠杆菌-枯草芽孢杆菌穿梭载体,从地衣芽孢杆菌ATCC 14580菌株基因组文库得到一个强度较高的启动子的克隆子P111509,克隆子P111509在大肠杆菌DH5α中所表达的β-半乳糖苷酶酶活有1290 miller单位,在枯草杆菌1A747中有3332 miller单位.将P111509基因启动子片断测序后与地衣芽孢杆菌ATCC 14580基因组序列进行同源性分析,二者同源性达到100%,且具有地衣芽孢杆菌ATCC 14580基因启动子的保守序列.

An efficient promoter P111509was separated from Bacillus licheniformis ATCC 14580 strain genomic library with E. coli-Bacillus subtilis shuttle vector by using the method of gene-targeting. Using the β-galactosidase gene as reporter gene, this efficient promoter has 1290 miller units in the E. coli DHSa strain and 3332 miller units in Bacillus subtilis 1A747 strain. Sequencing the efficient promoter P111509 nucleotide sequence and comparing it with Bacillus Licheniformis ATCC 14580 strain genome DNA sequence, it is obtained that their homology reaches 100% and P111509comes correctly from Bacillus Licheniformis ATCC 14580 and has the promoter conserved sequence .