[Cloning and analysis of promoter-active fragments from Corynebacterium glutamicum 10147].
OBJECTIVE
To clone promoter-active fragments from Corynebacterium glutamicum for further construction of expression vectors.


METHODS
Random Sau3A I digested fragments of C. glutamicum 10147 chromosome were shot-gun cloned into the promoter-probe vector pAKC6 and promoter activity of the inserted fragments was selected by chloramphenicol resistance of transformed C. glutamicum cells.


RESULTS
Thirty promoter-carrying fragments were isolated. Three C. glutamicum clones harboring pAKC6 with promoter fragments displayed chloramphenicol acetyltransferase (CAT) activity of more than 24 U/mg. The fragment F57 led to the highest CAT activity of 32.50 U/mg, even more than that produced by the promoter Ptrc, 26.33 U/mg.


CONCLUSIONS
The strength of promoter on fragments F21, F54 and F57 is as strong as promoter Ptrc in C. glutamicum. These fragments can be used to construct expression vector.
Liu
Guiming
G
Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China. guimingl@126.com
Zhao
Zhi
Z
Zhang
Yingzi
Y
Wang
Yu
Y
Ding
Jiuyuan
J
D001483:Base Sequence; D003001:Cloning, Molecular; D048230:Corynebacterium glutamicum / Q000235:genetics*; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic