Cloning and analysis of promoter-active fragments from Corynebacterium glutamicum 10147
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    Abstract:

    Abstract: [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. [Conclusion] 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.

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Guiming Liu, Zhi Zhao, Yingzi Zhang, Yu Wang, Jiuyuan Ding. Cloning and analysis of promoter-active fragments from Corynebacterium glutamicum 10147. [J]. Acta Microbiologica Sinica, 2009, 49(7): 972-977

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  • Received:January 05,2009
  • Revised:April 03,2009
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