CAJ | 학술논문

克隆组装了大肠杆菌的乙酰-CoA羧化酶基因acc,以研究其异源表达对变铅青链霉菌中2种“沉默”抗生素放线紫红素和十一烷基灵菌红素合成的影响。大肠杆菌ACC由4个基因编码,通过PCR扩增4个基因,并通过重叠延伸PCR加上ermE启动子,构建得到4个重组基因；再将重组基因依次组装得到异源表达质粒pHLZ11,接合转移到变铅青链霉菌中进行异源表达。结果显示：含有异源表达质粒的变铅青链霉菌接合转移子能合成放线紫红素而十一烷基灵菌红素产量提高。表明大肠杆菌acc基因异源表达能够激活变铅青链霉菌沉默抗生素的合成,并提高已有抗生素产量。
극륭조장료대장간균적을선-CoA최화매기인acc,이연구기이원표체대변연청련매균중2충“침묵”항생소방선자홍소화십일완기령균홍소합성적영향。대장간균ACC유4개기인편마,통과PCR확증4개기인,병통과중첩연신PCR가상ermE계동자,구건득도4개중조기인；재장중조기인의차조장득도이원표체질립pHLZ11,접합전이도변연청련매균중진행이원표체。결과현시：함유이원표체질립적변연청련매균접합전이자능합성방선자홍소이십일완기령균홍소산량제고。표명대장간균acc기인이원표체능구격활변연청련매균침묵항생소적합성,병제고이유항생소산량。
The Streptomyces lividans genome carries two gene clusters encoding the biosynthetic pathways of actinorhodin and undecylprodigiosin. However neither antibiotic is produced in S. lividans under the conditions of common laboratory growth. Malonyl-CoA as precursor was needed in the biosynthesis of both compounds catalyzed by the committed enzyme acetyl-CoA carboxylase ( ACC) . This study was aimed to clone the acc genes from Esch-erichia coli and introduce them into S. lividans to investigate their effect on the production of actinorhodin and un-decylprodigiosin. The E. coli ACC subunits are encoded by 4 acc genes. These acc genes were cloned with adding ermE promoter to each through overlapping extension PCR to gain 4 recombined genes. The recombinant genes were then cloned into pSET152 successively to gain a heterologous expression plasmid. The constructed plasmid was transferred into S. lividans by conjugation,and the antibiotic production in exconjugants was examined. The exconjugants could not only produce actinorhodin but also showed a prominent increase in the production of unde-cylprodigiosin. acc from E. coli can improve the production of pre-existing antibiotics and activate the expression of silent gene clusters as well in S. lividans.