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Generic Protocol for Optimization of Heterologous Protein Production Using Automated Microbioreactor Technology
Published on: December 15, 2017
Construction of genetic parts from the Corynebacterium glutamicum genome with high expression activities
Zihao Zhao1,2, Xiuxia Liu3,4, Wei Zhang1,2
1National Engineering Laboratory for Cereal Fermentation Technology, Jiangnan University, Wuxi, 214122, China.
Objective:
To construct effective genetic expression parts controlling transcription and translation initiation for synthetic biology and heterologous expression in Corynebacterium glutamicum.
Result:
Twelve highly expressed genes were identified from the proteomic data of C. glutamicum. Their related sequences were used to construct bicistronic genetic expression parts. Each part contain promoter, 5'-UTR, N-terminal sequence of the source gene and a conserved SD sequence, associated with target gene, forming the bicistronic expression cassette. The enhanced green fluorescent protein (EGFP) expression levels controlled by these novel parts have 1.4 to 790-fold increase in C. glutamicum compared with corresponding promoter-5'-UTR part. One of the bicistronic parts is 1.35 times the EGFP expression of the constitutive-expression pXMJ19. These bicistronic parts had expression advantage compared with conventional promoter-5'-UTR parts.
Conclusion:
Various genetic parts for efficient gene expression can be quickly obtained via this new method.
