斑贝链霉菌接合转移系统的构建及透明颤菌血红蛋白基因的表达对其次级代谢的影响

Construction of Conjugal Transfer System of Streptomyces bambergiensis and the Effects of Expression of Vitreoscilla Hemoglobin on its Secondary Metabolites

以pSET152和pHZ1358为出发质粒,通过供体大肠杆菌ET12567(pUZ8002)和S17-1接合转移斑贝链霉菌(Streptomycesbambergiensis),构建和优化了接合转移体系。采用OOE-PCR技术构建含ermEp*与vhb结构基因融合片段的整合型表达载体pBIB2005,转化ET12567(pUZ8002)后,属间接合转移至斑贝链霉菌。通过PCR、CO结合差光谱验证vhb基因在斑贝链霉菌中整合表达。摇瓶发酵显示VHb蛋白能改善菌体对氧的需求,一定程度上促进细胞生长,提高抗生素产量。

Intergeneric transfer of plasmid vectors pSET152 and pHZ1358 from Donor E. coli ET12567 （pUZ8002） and S17-1 to Streptomyees bambergiensis was demonstrated and optimized . Integrating plasmid pBIB2005 containing ermEp^＊ and construction gene of vhb, constructed by OOE-PCR , was transformed to ET12567 （ pUZ8002 ）, and then transfered to Streptomyces bambergiensis by conjugation. The integrating expression of vhb gene in Streptomyces bambergiensis was verified by PCR and CO binding difference spectrum. The fermentation results showed VHb could improve oxygen demand, promote cell growth and enhance antibiotics production.