Construction of genetic parts from the Corynebacterium glutamicum genome with high expression activities

Biotechnol Lett. 2016 Dec;38(12):2119-2126. doi: 10.1007/s10529-016-2196-y. Epub 2016 Aug 31.

Abstract

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.

Keywords: Corynebacterium glutamicum; Gene expression; Genetic part; Proteomics; Translation coupling.

MeSH terms

  • Corynebacterium glutamicum / genetics*
  • Corynebacterium glutamicum / metabolism
  • Genome, Bacterial / genetics*
  • Proteomics / methods