Codon-optimized DsRed fluorescent protein for use in Mycobacterium tuberculosis.

Area of Science:

• Microbiology
• Molecular Biology
• Biochemistry

Background:

• Mycobacterium tuberculosis research requires robust fluorescent protein tools.
• Previous work codon-optimized mCherry, tdTomato, and Turbo-635 for mycobacteria.

Purpose of the Study:

• To expand the available repertoire of red fluorescent proteins for Mycobacterium tuberculosis.
• To codon-optimize and characterize DsRed for expression in mycobacteria.

Main Methods:

• Generated expression constructs for full-length DsRed using strong constitutive promoters (P_hsp60, P_rpsA, P_G13).
• Confirmed expression and fluorescence of full-length DsRed in mycobacteria.
• Assessed functionality of truncated DsRed variants.

Main Results:

• Full-length DsRed was successfully expressed and exhibited bright fluorescence in mycobacteria.
• Truncated DsRed variants lacking N-terminal amino acids were non-functional.
• The codon-optimized DsRed expands the toolkit for Mycobacterium tuberculosis research.

Conclusions:

• Codon-optimized full-length DsRed is a functional and bright red fluorescent protein for Mycobacterium tuberculosis.
• This optimization enhances imaging and reporter applications in tuberculosis studies.
• The expanded repertoire of fluorescent proteins aids in understanding mycobacterial biology.