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Design of Ribosome Binding Sites in Streptomyces coelicolor

[ Vol. 14 , Issue. 4 ]

Author(s):

Hong-Dou Luo, Yang Tao, Wen-Guang Wang, Tao Lin, Yue-Yue Wang* and Hui Jiang*   Pages 287 - 292 ( 6 )

Abstract:


Background: The ribosome binding site (RBS) containing Shine-Dalgarno (SD) sequence in prokaryotes plays an important role in identification of the translation initiation site within mRNA by ribosome.

Objective: Our study aimed to improve the target protein production under translational level in prokaryotes by engineering of RBSs.

Method: Two RBSs were designed in Streptomyces coelicolor M145, Sco-RBS* with an SD sequence which is completely complementary to 3'-end of 16S RNA and Sco-RBS0 with an SD sequence which is completely non-complementary to 3'-end of 16S RNA. The enhanced green fluorescent protein (EGFP) was used as a reporter, whose gene was transcribed and translated under the control of Sco- RBS*, Sco-RBS0, and a native RBS Sco-RBSactI in recombinant S. coelicolor strains.

Results: Replacement of Sco-RBSactI with Sco-RBS* resulted in increase of both EGFP production and the ratio of EGFP to EGFP mRNA by 2.67-fold and 6.07-fold, respectively. Replacement of Sco- RBSactI with Sco-RBS0 resulted in decrease of both EGFP and the ratio of EGFP to EGFP mRNA by 4.35-fold and 2.18-fold, respectively.

Conclusion: We provided a method to increase protein production at the translational level in Streptomyces.

Keywords:

Binding site, EGFP production, enhanced green fluorescent protein, recombinant strains, ribosome, Streptomyces coelicolor.

Affiliation:

College of Life Sciences, Zhejiang University, Hangzhou, Zhejiang, 310058, College of Life Sciences, Zhejiang University, Hangzhou, Zhejiang, 310058, College of Life Sciences, Zhejiang University, Hangzhou, Zhejiang, 310058, Nanjing Enzymetree Biotechnology Co., Ltd. Nanjing, Jiangsu, 211100, School of Life Sciences, Fudan University, Shanghai, 200438, College of Life Sciences, Zhejiang University, Hangzhou, Zhejiang, 310058

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