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Significant expression of a Chinese scorpion peptide, BmK1, in Escherichia coli through promoter engineering and gene dosage strategy.

Wang J, Xiong Z, Yang Y, Zhao N, Wang Y - Biotechnol. Appl. Biochem. (2013)

Bottom Line: Heterologous expression is an efficient alternative to conventional extraction to produce a specific Buthus martensii Karsch (BmK) peptide.The yield of the purified BmK1 achieved 196.74 mg L(-1) in E. coli BL21(DE3) pJF431, which was improved 2.09-fold compared with the control.This was the highest reported production of scorpion peptides in E. coli.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, People's Republic of China; State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, People's Republic of China.

No MeSH data available.


Related in: MedlinePlus

Effect of gene dosage on BmK1 expression. (A) Multicopy expression cassette construction. (B) Effect of gene dosage on cell growth. (C) Effect of gene dosage on BmK1 yield.
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fig05: Effect of gene dosage on BmK1 expression. (A) Multicopy expression cassette construction. (B) Effect of gene dosage on cell growth. (C) Effect of gene dosage on BmK1 yield.

Mentions: When the best promoter strength was confirmed for BmK1 expression, gene dosage strategy was used to further increase the expression of BmK1. One, two, three, and four copies of BmK1 gene expression cassettes were assembled under the control of the best promoter (Fig.5A). The expression level of BmK1 with two, three, and four copies controlled by the promoter PpJF325 significantly increased along with a slight growth inhibition during the whole process (Figs.5B and 5C). The maximum BmK1 content of pJF430, pJF431, and pJF432 reached 20.6%, 21.79%, and 20.73%, respectively. The maximum BmK1 content using the optimal promoter strength and gene copy number (pJF431) was improved 3.56-fold compared with that of the control (pJF391) using the trc promoter and a single copy of gene, suggesting that it is a simple and efficient method to express BmK1 through promoter engineering and gene dosage strategy.


Significant expression of a Chinese scorpion peptide, BmK1, in Escherichia coli through promoter engineering and gene dosage strategy.

Wang J, Xiong Z, Yang Y, Zhao N, Wang Y - Biotechnol. Appl. Biochem. (2013)

Effect of gene dosage on BmK1 expression. (A) Multicopy expression cassette construction. (B) Effect of gene dosage on cell growth. (C) Effect of gene dosage on BmK1 yield.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4269186&req=5

fig05: Effect of gene dosage on BmK1 expression. (A) Multicopy expression cassette construction. (B) Effect of gene dosage on cell growth. (C) Effect of gene dosage on BmK1 yield.
Mentions: When the best promoter strength was confirmed for BmK1 expression, gene dosage strategy was used to further increase the expression of BmK1. One, two, three, and four copies of BmK1 gene expression cassettes were assembled under the control of the best promoter (Fig.5A). The expression level of BmK1 with two, three, and four copies controlled by the promoter PpJF325 significantly increased along with a slight growth inhibition during the whole process (Figs.5B and 5C). The maximum BmK1 content of pJF430, pJF431, and pJF432 reached 20.6%, 21.79%, and 20.73%, respectively. The maximum BmK1 content using the optimal promoter strength and gene copy number (pJF431) was improved 3.56-fold compared with that of the control (pJF391) using the trc promoter and a single copy of gene, suggesting that it is a simple and efficient method to express BmK1 through promoter engineering and gene dosage strategy.

Bottom Line: Heterologous expression is an efficient alternative to conventional extraction to produce a specific Buthus martensii Karsch (BmK) peptide.The yield of the purified BmK1 achieved 196.74 mg L(-1) in E. coli BL21(DE3) pJF431, which was improved 2.09-fold compared with the control.This was the highest reported production of scorpion peptides in E. coli.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, People's Republic of China; State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, People's Republic of China.

No MeSH data available.


Related in: MedlinePlus