Related Experiment Video
Updated: Jul 26, 2026
13:28
Single-cell Analysis of Bacillus subtilis Biofilms Using Fluorescence Microscopy and Flow Cytometry
Published on: February 15, 2012
Genome-scale expression of proteins from Bacillus subtilis
S Moy1, L Dieckman, M Schiffer
1Biosciences Division, Argonne National Laboratory, Argonne, IL 60439, USA.
Journal of Structural and Functional Genomics
|July 21, 2004
Summary
High-throughput cloning and expression methods were developed for over 850 Bacillus subtilis genes. This automated process successfully produced soluble proteins for structural genomics, showing promise for future target discovery.
Area of Science:
- Genomics
- Structural Biology
- Biotechnology
Background:
- Structural genomics programs require efficient methods for gene cloning and protein expression.
- Bacillus subtilis is a model organism with a well-characterized genome, making it a suitable target for large-scale studies.
Purpose of the Study:
- To develop and evaluate high-throughput methods for cloning and expressing Bacillus subtilis genes.
- To assess the feasibility of plate-based methods for various protein types (cytoplasmic, secretory, membrane domains).
- To generate soluble protein clones for structural determination pipelines.
Main Methods:
- Application of automated, 96-well plate-based methods for primer design, cloning, and expression.
- Utilized tag detection screens to identify soluble protein production.
- Evaluated methods for cytoplasmic, secretory, and membrane protein domains.
Main Results:
- Successfully cloned and expressed over 850 Bacillus subtilis genes.
- Produced soluble protein clones suitable for structural genomics.
- Demonstrated the feasibility of plate-based cloning and expression for diverse protein targets.
Conclusions:
- Automated, high-throughput cloning and expression methods are effective for generating protein resources for structural genomics.
- These plate-based approaches show promise for expanding target availability for structure determination.
- Continued technological development in automation is crucial for advancing structural genomics programs.
More Related Videos
06:45Monitoring Intraspecies Competition in a Bacterial Cell Population by Cocultivation of Fluorescently Labelled Strains
Published on: January 18, 2014
12:23Recombinant Protein Expression, Crystallization, and Biophysical Studies of a Bacillus-conserved Nucleotide Pyrophosphorylase, BcMazG
Published on: May 16, 2017
Related Concept Videos
00:43Bacterial RNA Polymerase
Unlike eukaryotes, bacteria use a single RNA Polymerase (RNAP) to transcribe all genes. The different subunits of bacterial RNAPhave distinct functions. The multisubunit structure of the bacterial RNAP helps the enzyme to maintain catalytic function, facilitate assembly, interact with DNA and RNA, and self-regulate its activity.
In most genes, the transcription site is a single base present upstream of the coding sequence. Though RNAP is a catalytically efficient enzyme, it does not recognize...
In most genes, the transcription site is a single base present upstream of the coding sequence. Though RNAP is a catalytically efficient enzyme, it does not recognize...
01:29Coordination of Gene Expression Processes in Bacteria
The DNA replication, transcription, and translation processes are intricately coupled in bacteria, allowing efficient gene expression and rapid protein synthesis. While this physical and functional coordination is advantageous, it introduces challenges that bacteria overcome through specific regulatory mechanisms.Coupling of Replication, Transcription, and TranslationThe coupling of replication, transcription, and translation is a hallmark of bacterial gene expression. As the replisome unwinds...
01:26Bacterial Translocation and Protein Secretion
Bacterial protein secretion involves translocation systems to ensure proteins reach their designated locations, including the plasma membrane, periplasm, outer membrane, or the external environment. These translocation systems are vital for bacterial physiology, supporting processes like membrane assembly, enzymatic activity in the periplasm, and interactions with the external environment. The division of labor between Sec and Tat pathways ensures efficiency in handling proteins with diverse...
01:28Global Regulatory Systems
Global regulatory systems in bacteria enable rapid and coordinated responses to environmental changes by integrating sensory inputs with gene expression, ensuring efficient adaptation to fluctuating conditions. Key global regulatory mechanisms include regulons, two-component systems, sigma factors, and secondary messengers.Regulons and Global RegulatorsA regulon is a collection of genes and operons controlled by a common global regulator. These regulators enable bacteria to prioritize resource...
01:17Gene Regulation During Sporulation
Sporulation is a complex developmental process that allows certain Gram-positive bacteria, such as Bacillus subtilis and Clostridium species, to survive extreme environmental conditions. This process is tightly regulated by a series of signaling cascades and transcriptional controls, ensuring the formation of a highly resistant endospore.Sporulation is triggered by unfavorable conditions, such as nutrient depletion, and is governed by a phosphorelay system. One of the sensor kinases, such as...
01:23Stringent Response in E. coli
Bacterial growth is closely tied to nutrient availability, with cells proliferating exponentially under favorable conditions and entering a stationary phase when resources become scarce. This transition is mediated by a regulatory mechanism known as the stringent response, which allows bacteria to adapt to nutrient deprivation by modulating gene expression and metabolic activity.During nutrient scarcity, intracellular amino acid levels decline. It results in the accumulation of uncharged tRNAs...