Title Escherichia coli와 corynebacterium glutamicum간의 shuttle vectors의 C. glutamicum에서의 안정성에 대한 클론된 유전자의 영향
Author 노갑수 · * 김성준 · 오종원 · 이현환 · 현형환 · * 이재홍
Address 제일제당(주) 종합연구소 · * 제일제당(주) 가공식품개발센터
Bibliography Korean Journal of Microbiology, 29(3),149-154, 1991
Key Words Plasmid stability, Corynebacterium glutamicum, Shuttle vector
Abstract Escherichia coli/Corynebacterium glutamicum shuttle vectors, pECCG1 and pECCG2 were constructed by joining a 3.00 kb cryptic plasmid pCB 1 from C. glutamicum and a 3.94 kb plasmid pACYC 177 from E. coli. By trimming unessential parts and introducing mulitiple cloning site into the plasmid pECCG 1, a plasmid pECCG122(5.1kb) was constructed. All the shuttle vectors were stably maintained in C. glutamicum up to about 40 generations irrespective of kanamycin addition in the medium. Threonine operon (homoserine dehydrogenase/homoserine kinase) and dapA gene (dihydrodipicolinate synthetase) of C. glutamicum were cloned into the plasmid pECCG122, and the resultant plasmids were designated pTN31 and pDHDP19, respectively. They were used to study the effect of cloned foreign gene on the stability of the plasmid pECCG122. Plasmids pTN31 and pDHDP19 were segregated rapidly from C. glutamicum when cultured in the medium without kanamycin. In medium with 50㎍/㎖ of kanamycin, their segregation rates were much slower than those in medium without kanamycin, but the danamycin addition didn't guarantee the complete maintenance of the plasmids in C. glutamicum.
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