Selective Capture of Transcribed Sequences: A Promising Approach for Investigating Bacterium-Insect Interactions.

Ruisheng An,Parwinder S Grewal

doi:10.3390/insects3010295

Ruisheng An, Parwinder S Grewal

Open Access

https://doi.org/10.3390/insects3010295

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Journal: Insects	Publication Date: Mar 2, 2012
Citations: 46	License type: CC BY 4.0

Affiliation: The Ohio State University

Abstract

Bacterial interactions with eukaryotic hosts are complex processes which vary from pathogenic to mutualistic. Identification of bacterial genes differentially expressed in the host, promises to unravel molecular mechanisms driving and maintaining such interactions. Several techniques have been developed in the past 20 years to investigate bacterial gene expression within their hosts. The most commonly used techniques include in-vivo expression technology, signature-tagged mutagenesis, differential fluorescence induction, and cDNA microarrays. However, the limitations of these techniques in analyzing bacterial in-vivo gene expression indicate the need to develop alternative tools. With many advantages over the other methods for analyzing bacterial in-vivo gene expression, selective capture of transcribed sequences (SCOTS) technique has the prospect of becoming an elegant tool for discovery of genes involved in the bacterium-host interaction. Here, we summarize the advances in SCOTS technique, including its current and potential applications in bacterial gene expression studies under a variety of conditions from in-vitro to in-vivo and from mammals to insects.

Highlights

The interactions between bacteria and their eukaryotic hosts require the activity of many gene products
The translational-level methods mainly refer to two-dimensional gel electrophoresis (2-D gel) and multidimensional protein identification technology (MUDPIT)
While the rest of the techniques have been originally developed to study the bacterial gene expression in mammalian models, some of them have been subsequently applied in bacteriainsect interactions

Summary

Introduction

The interactions between bacteria and their eukaryotic hosts require the activity of many gene products. Multiple techniques have become available to scan bacterial genes preferentially expressed under a specific growth condition. These techniques are based on determination of either differentially expressed bacterial genes (transcriptional level) or produced bacterial proteins (translational level). PCR-hybridization based methods include differential-display reverse transcription-PCR (DDRT-PCR), cDNA microarray, and selective capture of transcribed sequences (SCOTS). Among these techniques, 2-D gel, MUDPIT, EGT and DDRT-PCR have been applied for profiling bacterial genes expressed under in-vitro conditions, but not in-vivo. The main purpose of this paper is to share updated information about the techniques and to describe the proper conditions for the use of each technique the SCOTS technique

In-vitro Profiling Techniques

Mutagenesis Based Methods

Hybridization Based Methods

Selective Capture of Transcribed Sequences

SCOTS versus other Methods

Findings

Conclusions