Cloning and Identification of a Gene Cluster in Streptomyces Spectabilis-Spectinomycin Producer

다국어 초록 (Multilingual Abstract)

A biosynthetic gene cluster involved in the production of spectinomycin was cloned from Streptomyces spectabilis cosmid library with dTDP-glucose 4,6-dehydratase and aminotransferase gene probes. The DNA probes were obtained from S, spectabilis genome with primers based on consensus sequences of known enzymes using a polytmrase chain reaction. One of the cloned cosmids, pSPMB, contained the homologues of the dTDP-glucose 4,6-dchydratase and aminotransferase genes. The pSPM58 including a spectinomycin resistance homologous gene from pSPM8 cosmid was isolated and sequenced. The DNA sequence analysis revealed four open reading frames and one incomplete reading frame. The putative orfl protein reveals 29% amino acid sequence homology of putative myo-inositol-2-dehydrogenase from Vibrio anguillarnm. The orf2 is about 46% homologous to putative myo-inositol-1-monophosphotase of Mycobacterium leprae. The orf3 showed strong homology (67.4%) to putative export protein (spcT) from S. flavopersicus. The amino acid sequence deduced from orf4 showed a high similarity (70.8%) to a spectinomycin resistance gene (spcN) from S. favnpersicus. Expression of orf4 in E. coli BL21 showed the spectinomycin resistance.

목차 (Table of Contents)

• INTRODUCTION = 209
• MATERIALS AND METHODS == 214
• REFERENCES = 218