除虫链霉菌uhk01sav基因的功能研究微生物学专业论文.docxVIP

扬州大学硕士学位论文Functional 扬州大学硕士学位论文 Functional study of uhkOlsav gene in Streptomyces a1，e厂7即Z tt lts M．S．Candidate：Juanmei He Supervisor：Prof．Guoqiang Zhu；Associate Prof．Yinhua Lu Abstract Streptomyces is a genus of Gram．positive bacteria and possess strong secondary metabolism．It iS a prominent producer of a wide variety of antibiotics and other bioactive compounds．which are widely applied in human and veterinary medicine as well as agriculture．So far,researches on regulation of antibiotic biosynthesis in Streptomyces were mostly carded out in the model strain-．Streptomyces coelicolor．In OUlprevious study,we showed that deletion of“五^刀1 gene in．孓coelicolor M145．which encodes an unpaired histidine kinase(HK)，led to not only obviously increased biosynthesis of actinorhodin(Act)and undecylprodigiosin(Red)，but also defective development， including colony color from grey to red，formation of sparse aerial hypha and spores． BioinfoITnatics analysis shows that uhko i gene homologues are widely distributed in other different Streptomyces species．In the present study,functional study Was carried out to identify the“^忌口1 homologous gene uhkOlsav from all important industrial antibiotic producer-．Streptomyces avermitilis．We found that．similar as the function of uhk01 in coelicolor．“五尼口lsav alSO play important roles in the regulation of antibiotic biosynthesis and development inavermitilis． Inavermitilis NRRL8 1 65 strain．uhkOlsav-deletion mutant(8 1 65△uhkOlsav) constructed by PCR．targeting system showed increased biosynthesis of oligomycin A。 approximately 5 to 6．fold higher than that of the wild type．Moreover,the mutant grew slower and showed defective sporulation when compared with NRRL8 1 65．The increased synthesis of oligomycin A and defective sporulation could be readily rescued by complementation with“矗加1say gene，suggesting that uhkOlsav was a negative regulator for oligomycin biosynthesis and had a close relationship with sporulation．In order to lealTl more about the mole