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KMID : 0578319960060060712
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Molecules and Cells
1996 Volume.6 No. 6 p.712 ~ p.718
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Optimization of the Bovine Growth Hormone Gene Expression in E. coli
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Choi Jang-Won
Lee Se-Yong
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Abstract
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To achieve the high level expression of the bovine growth hormone (bGH) gene in E. coli, the 5´-coding region of the bGH cDNA was modified by replacing the natural sequence with various synthetic DNA fragments resulting in the change of mRNA secondary structure. The expression plasmids have been constructed using the pKK233-2 vector containing the trc promoter (pKBJl-14). All induced cells harboring these plagmids did not produce a detectable level of bGH. However, replacement of the replication origin with the mutated CoIEl replicon resulted in a substantial increase of the bGH production to 3.9%, 27.4%, and 10.9% of total cell protein in pUBJ7, pUBJ9, and pUBJ10, respectively. The analysis of transcripts by dot blot hybridization showed that a post-transcriptional control mechanism is a critical factor for the efficient expression of the bGH gene.
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