CAJ | 학술논문

提取了侧孢短芽孢杆菌X10的基因组DNA,以绿色荧光蛋白基因(green fluorescent protein,gfp)为报告基因,以启动子探针pUC19-GFP为载体,通过鸟枪法在大肠杆菌DH5α中构建了X10的启动子文库,通过筛选获得了14个阳性克隆,编号为P1～P14.测定了阳性克隆子的荧光强度,结果表明P6中gfp基因的启动子活性最强,它的荧光强度达到了355.67,而P14中gfp基因的启动子活性最弱,它的荧光强度只有211.67.对P6克隆中的重组质粒的插入片段进行了测序和序列分析.
제취료측포단아포간균X10적기인조DNA,이록색형광단백기인(green fluorescent protein,gfp)위보고기인,이계동자탐침pUC19-GFP위재체,통과조창법재대장간균DH5α중구건료X10적계동자문고,통과사선획득료14개양성극륭,편호위P1～P14.측정료양성극륭자적형광강도,결과표명P6중gfp기인적계동자활성최강,타적형광강도체도료355.67,이P14중gfp기인적계동자활성최약,타적형광강도지유211.67.대P6극륭중적중조질립적삽입편단진행료측서화서렬분석.
High quality genomic DNA of Brevibacillus laterosporus X10 was extracted. The promoter library of X10 was constructed in Escherichia coli DH5α with pUC19-CFP as promoter vector by shotgun-cloning method. 14 positive clones designed as P1～P14 were obtained by screening, the fluorescent intensity of which was assayed to compare the promoting strength of the promoters. The results showed that the promoter of gjp in P6 was the strongest with fluorescent intensity of 3SS. 67 ,while the promoter of gfp in P14 was the weakest with fluorescent intensity of 211.67. The inserting fragments in the recombinant from P6 were sequenced and analyzed.