Phenotypic Comparison of Wild-Type and Mutant Strains of Streptomyces coelicolor

Abstract

Source: Bennett, J. A., et al. Visual and Microscopic Evaluation of Streptomyces Developmental Mutants. J. Vis. Exp. (2018)

This video demonstrates the cultivation and phenotypic comparison of wild-type and mutant Streptomyces coelicolor strains. It outlines procedures for minimizing spontaneous mutations, preventing cross-contamination, and promoting bacterial growth through proper plating techniques. Finally, the video highlights visual differences in colony morphology to illustrate the developmental effects of mutations in the bacterial strains.

Protocol

1. Prepare Instruments, Culture Media, Solutions, and Petri Dishes

1. Autoclave toothpicks in a 50 mL beaker covered with aluminum foil to keep them sterile.

2. Sterilize all dry goods that will be used (tips, sticks, glassware, etc.) in an autoclave.

NOTE: Caution! Follow all safety directions for the autoclave model used. Autoclaves pose a risk for explosions, burns from steam, and contact with hot surfaces, materials, and media.

3. Prepare the MS agar (Mannitol Soya Flour (SFM)) to grow cultures.

Results

Figure 1: Photograph of a representative agar plate demonstrating macroscopic colony phenotypes of <.......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
Sterile Water	GeneMate	G-3250-1L
Toothpicks (flat)	Target	081-22-1957
Pipette Tips (10 mL)	GeneMate	P-1240-10
Pipette Tips (200 mL)	GeneMate	P-1240-200Y
Pipette Tips (1,250 mL)	GeneMate	P-1240-1250
Cotton Swabs	Fisherbrand	23-400-124
Soy Flour	Bob's Red Mill	1516C164
D-Mannitol	Sigma-Aldrich	M4125-1KG
Agar	Sigma-Aldrich	A1296-1KG
Glycerol 100%	VWR amresco life science	0854-1L
0.8% NaCl (Saline)	Sigma-Aldrich	SLBB9000V
Petri dishes	Sigma-Aldrich	P5856-500EA
Cover slips #1.5	Thomas Scientific	64-0721
Slides	Carolina	63-2010
Autoclave	Tuttnauer	2540E
Forceps	Carolina Biological	624504
Bunsen Burner	Carolina Biological	706706
Methanol	Sigma-Aldrich	34860-1L-R
PBS (phosphate buffer saline)	Sigma-Aldrich	P4417-50TAB
100 mL beaker	Pyrex USA	1000-T
1 L beaker	Carolina Biological	721253