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All Journal ANNALES BOGORIENSES Teknologi Indonesia
Handayani, Ira
Research Center for Biotechnology - Indonesian Institute of Sciences (LIPI)
Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Chemical Engineering, Chemistry & Bioengineering Chemistry Computer Science & IT Immunology & microbiology Medicine & Pharmacology

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Journal : Teknologi Indonesia

 1 
EXPRESSION OF THE FLUORESCENT PROTEIN MTURQUOISE2 IN THE PERIPLASM OF Escherichia coli Handayani, Ira; Kusharyoto, Wien
Teknologi Indonesia Vol 39, No 3 (2016)
Publisher : LIPI Press

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (16.054 KB) | DOI: 10.14203/jti.v39i3.288

Abstract

Many variants of cyan fluorescent protein (CFP) have been developed as fluorescent tags which are widely used as donors in Förster resonance energy transfer (FRET) experiments. Recent improvement of CFP variants resulted in mTurquoise2, a brighter variant with faster maturation, high photostability, longer mono-exponential lifetime and the highest quantum yield measured for a monomeric fluorescent protein. Here, the authors describe the expression of mTurquoise2 targeted for secretion via the general secretory (Sec) translocation pathway into the highly oxidizing periplasm of Escherichia coli. The use of signal peptide MPB*1, a modified signal sequence of maltose binding protein was investigated. The His6-tagged fluorescent protein was expressed in E. coli NiCo21(DE3) and purified by means of immobilized metal ion affinity chromatography (IMAC) on TALON™ matrix. In SDSPAGE and Western blot analysis, a single band corresponding to a molecular mass of approximately 28 kDa was observed, which correlated with the predicted molecular mass based on the amino acid sequence of mTurquoise2. 
Co-Authors Asrul Muhamad Fuad, Asrul Muhamad Dian Andriani, Dian Kusharyoto, Wien Kusharyoto, Wien Martha Sari