Optimization of DsRed production in Escherichia coli:: Effect of ribosome binding site sequestration on translation efficiency

被引：21

作者：

Pfleger, BF

Fawzi, NJ

Keasling, JD ^{[1
]}

机构：

[1] Univ Calif Berkeley, Dept Chem Engn, Berkeley, CA 94720 USA

[2] Univ Calif Berkeley, Dept Bioengn, Berkeley, CA USA

[3] Lawrence Berkeley Natl Lab, Dept Synthet Biol, Phys Biosci Div, Berkeley, CA USA

来源：

BIOTECHNOLOGY AND BIOENGINEERING | 2005年 / 92卷 / 05期

关键词：

D O I：

10.1002/bit.20630

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

DsRed-Express, a popular reporter protein, cannot be expressed in Escherichia coli using a consensus ribosome binding site (RBS) potentially due to base-pairing in the RBS that inhibits translation initiation. Saturation mutagenesis was used to probe for a gene sequence that minimized base-pairing in the RBS while maintaining the same spectral properties and maturation characteristics as DsRed-Express. The new DsRed, designated here as RFPEC for E. coli optimized red fluorescent protein, fluoresces 2.5 times greater than DsRed-Express when expressed from the same vector. (c) 2005 Wiley Periodicals, Inc.

引用

页码：553 / 558

页数：6

共 17 条

[1] Rapidly maturing variants of the Discosoma red fluorescent protein (DsRed) [J].