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Updated: Jun 19, 2026
Methods to Discover Alternative Promoter Usage and Transcriptional Regulation of Murine Bcrp1
Published on: May 27, 2016
[Cloning and analysis of promoter-active fragments from Corynebacterium glutamicum 10147]
Guiming Liu1, Zhi Zhao, Yingzi Zhang
1Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China. guimingl@126.com
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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