Abstract:
:Recently developed techniques allow genomic DNA sequencing from single microbial cells [Lasken RS: Single-cell genomic sequencing using multiple displacement amplification. Curr Opin Microbiol 2007, 10:510-516]. Here, we focus on research strategies for putting these methods into practice in the laboratory setting. An immediate consequence of single-cell sequencing is that it provides an alternative to culturing organisms as a prerequisite for genomic sequencing. The microgram amounts of DNA required as template are amplified from a single bacterium by a method called multiple displacement amplification (MDA) avoiding the need to grow cells. The ability to sequence DNA from individual cells will likely have an immense impact on microbiology considering the vast numbers of novel organisms, which have been inaccessible unless culture-independent methods could be used. However, special approaches have been necessary to work with amplified DNA. MDA may not recover the entire genome from the single copy present in most bacteria. Also, some sequence rearrangements can occur during the DNA amplification reaction. Over the past two years many research groups have begun to use MDA, and some practical approaches to single-cell sequencing have been developed. We review the consensus that is emerging on optimum methods, reliability of amplified template, and the proper interpretation of 'composite' genomes which result from the necessity of combining data from several single-cell MDA reactions in order to complete the assembly. Preferred laboratory methods are considered on the basis of experience at several large sequencing centers where >70% of genomes are now often recovered from single cells. Methods are reviewed for preparation of bacterial fractions from environmental samples, single-cell isolation, DNA amplification by MDA, and DNA sequencing.
journal_name
Curr Opin Microbioljournal_title
Current opinion in microbiologyauthors
Ishoey T,Woyke T,Stepanauskas R,Novotny M,Lasken RSdoi
10.1016/j.mib.2008.05.006subject
Has Abstractpub_date
2008-06-01 00:00:00pages
198-204issue
3eissn
1369-5274issn
1879-0364pii
S1369-5274(08)00063-5journal_volume
11pub_type
杂志文章,评审abstract::The obligatory intracellular bacterial pathogens Anaplasma and Ehrlichia infect leukocytes by hijacking host-cell components and processes. The type IV secretion system is up-regulated during infection. Among type IV secretion candidate substrates, an ankyrin repeat protein of Anaplasma phagocytophilum, AnkA, is deliv...
journal_title:Current opinion in microbiology
pub_type: 杂志文章,评审
doi:10.1016/j.mib.2009.12.008
更新日期:2010-02-01 00:00:00
abstract::Biological nitrogen fixation is a much more important process in the nitrogen cycle of the oceans than previously thought. Further, nitrogen fixation may have an influence on the capacity of the oceans to sequester carbon. A greater diversity of marine nitrogen fixers has also been uncovered but their quantitative sig...
journal_title:Current opinion in microbiology
pub_type: 杂志文章
doi:10.1016/s1369-5274(00)00215-0
更新日期:2001-06-01 00:00:00
abstract::Microbial communities have considerable impacts on animal health. However, only in recent years have the host factors impacting microbiome composition been explored. An increasing wealth of microbiome data in combination with decades of research on behavior, physiology, and development have resulted in the European ho...
journal_title:Current opinion in microbiology
pub_type: 杂志文章,评审
doi:10.1016/j.mib.2019.08.001
更新日期:2019-08-01 00:00:00
abstract::Elimination of non-functional or unwanted proteins is critical for cell growth and regulation. In bacteria, ATP-dependent proteases target cytoplasmic proteins for degradation, contributing to both protein quality control and regulation of specific proteins, thus playing roles parallel to that of the proteasome in euk...
journal_title:Current opinion in microbiology
pub_type: 杂志文章,评审
doi:10.1016/j.mib.2013.01.002
更新日期:2013-04-01 00:00:00
abstract::In this paper, we discuss recent advances in our knowledge of the dengue virus life cycle based on new structural data of the virus and its proteins. Specifically, we focus on the structure of the pre-membrane protein, prM and its role in virus assembly, the first full-length structure of a multi-domain dengue virus r...
journal_title:Current opinion in microbiology
pub_type: 杂志文章,评审
doi:10.1016/j.mib.2008.06.004
更新日期:2008-08-01 00:00:00
abstract::Candida albicans is a regular member of the intestinal microbiota in the majority of the human population. This underscores C. albicans' adaptation to life in the intestine without inducing competitive interactions with other microbes, or immune responses detrimental to its survival. However, specific conditions such ...
journal_title:Current opinion in microbiology
pub_type: 杂志文章,评审
doi:10.1016/j.mib.2020.05.006
更新日期:2020-08-01 00:00:00
abstract::Viruses attach to specific receptors, but identified receptors are often absent from the epithelial surfaces where infections begin. Viruses can bind to alternative receptor molecules present on epithelial surfaces and they can enter hosts in ways that bypass mucosal barriers to infection. To understand how viruses cr...
journal_title:Current opinion in microbiology
pub_type: 杂志文章,评审
doi:10.1016/s1369-5274(03)00097-3
更新日期:2003-08-01 00:00:00
abstract::Haloalkaliphiles are double extremophilic organisms thriving both at high salinity and alkaline pH. Although numerous haloalkaliphilic representatives have been identified among Archaea and Bacteria over the past 15 years, the adaptations underlying their prosperity at haloalkaline conditions are scarcely known. A mul...
journal_title:Current opinion in microbiology
pub_type: 杂志文章,评审
doi:10.1016/j.mib.2015.05.003
更新日期:2015-06-01 00:00:00
abstract::In the past year, our knowledge of type IV transporters of Gram-negative bacteria has further expanded. Advances include the discovery of additional members of this family of proteins, increased knowledge of the morphologies of type IV transporters, and a better understanding of the mechanisms by which macromolecules ...
journal_title:Current opinion in microbiology
pub_type: 杂志文章,评审
doi:10.1016/s1369-5274(99)80004-6
更新日期:1999-02-01 00:00:00
abstract::Bacterium-host interactions are important for diverse ecological settings including pathogenicity and symbiosis. Electron cryotomography is a powerful method for studying the macromolecular complexes that mediate such interactions in situ. The main limitation of electron cryotomography is its restriction to relatively...
journal_title:Current opinion in microbiology
pub_type: 杂志文章,评审
doi:10.1016/j.mib.2017.12.006
更新日期:2018-06-01 00:00:00
abstract::A combination of structural, biochemical and functional studies with the flavivirus tick-borne encephalitis virus has revealed the characteristics of a new class of viral fusion protein, class II, that is unrelated to the class I viral fusion proteins for which influenza virus hemagglutinin is the prototype. New struc...
journal_title:Current opinion in microbiology
pub_type: 杂志文章,评审
doi:10.1016/s1369-5274(00)00234-4
更新日期:2001-08-01 00:00:00
abstract::Clustered, regularly interspaced, short, palindromic repeats (CRISPR) loci, together with their CRISPR-associated (Cas) proteins, provide bacteria and archaea with adaptive immunity against invasion by bacteriophages, plasmids, and other mobile genetic elements. These host defenses impart selective pressure on phages ...
journal_title:Current opinion in microbiology
pub_type: 杂志文章,评审
doi:10.1016/j.mib.2017.06.003
更新日期:2017-06-01 00:00:00
abstract::The review highlights the dual role that the conserved master regulator FoxO has in aging by coordinating both stem cell proliferation and antimicrobial peptides, effector molecules of the innate immune system. As observed in non-senescent Hydra, continuously high activity of the transcription factor FoxO contributes ...
journal_title:Current opinion in microbiology
pub_type: 杂志文章,评审
doi:10.1016/j.mib.2017.06.009
更新日期:2017-08-01 00:00:00
abstract::In Gram-negative bacteria such as Escherichia coli the peptidoglycan sacculus resides in the periplasm, a compartment that experiences changes in pH value, osmolality, ion strength and other parameters depending on the cell's environment. Hence, the cell needs robust peptidoglycan growth mechanisms to grow and divide ...
journal_title:Current opinion in microbiology
pub_type: 杂志文章,评审
doi:10.1016/j.mib.2017.01.006
更新日期:2017-04-01 00:00:00
abstract::Natural hypersaline waters are widely distributed around the globe, as both continental surface waters and sea floor lakes, the latter being maintained by the large density difference between the hypersaline and overlying marine water. Owing to the extreme salt concentrations, close to or at saturation (approximately ...
journal_title:Current opinion in microbiology
pub_type: 杂志文章,评审
doi:10.1016/j.mib.2007.05.017
更新日期:2007-08-01 00:00:00
abstract::An increasingly diverse array of clinically relevant animal models of candidiasis have been established that mimic both the immune perturbations of the host and tissue-specific features of candidiasis in humans. Cause-and-effect analysis of Candida host-pathogen interactions using these animal models has made a quantu...
journal_title:Current opinion in microbiology
pub_type: 杂志文章,评审
doi:10.1016/j.mib.2004.06.001
更新日期:2004-08-01 00:00:00
abstract::Bacteria reside in externally accessible niches on and in multicellular organisms, often forming mutualistic relationships with their host. Recent studies have linked the composition of these microbial communities with alterations in the host's health, behavior, and development, yet the causative mediators of host-mic...
journal_title:Current opinion in microbiology
pub_type: 杂志文章,评审
doi:10.1016/j.mib.2017.05.006
更新日期:2017-08-01 00:00:00
abstract::RNA sequencing has emerged as the premier approach to study bacterial transcriptomes. While the earliest published studies analyzed the data qualitatively, the data are readily digitized and lend themselves to quantitative analysis. High-resolution RNA sequence (RNA-seq) data allows transcriptional features (promoters...
journal_title:Current opinion in microbiology
pub_type: 杂志文章,评审
doi:10.1016/j.mib.2014.11.011
更新日期:2015-02-01 00:00:00
abstract::Xenogeneic silencing proteins facilitate horizontal gene transfer by silencing expression of AT-rich sequences. By virtue of their activity these proteins serve as master regulators of a variety of important functions including motility, drug resistance, and virulence. Three families of silencers have been identified ...
journal_title:Current opinion in microbiology
pub_type: 杂志文章,评审
doi:10.1016/j.mib.2011.12.014
更新日期:2012-04-01 00:00:00
abstract::Associations between marine invertebrates and their cooperative bacterial symbionts offer access to an understanding of the roots of host-microbe interaction; for example, several symbioses like the squid-vibrio light organ association serve as models for investigating how each partner affects the developmental biolog...
journal_title:Current opinion in microbiology
pub_type: 杂志文章,评审
doi:10.1016/s1369-5274(00)00147-8
更新日期:2000-12-01 00:00:00
abstract::Host social interactions can provide multiple complex pathways for microbial transmission. Here, we suggest non-human primates as models to study the social transmission of commensal or mutualistic microbes due to their high sociality, wide range of group compositions and dominance structures, and diverse group intera...
journal_title:Current opinion in microbiology
pub_type: 杂志文章,评审
doi:10.1016/j.mib.2019.09.001
更新日期:2019-08-01 00:00:00
abstract::Sensing its changing environment is key for Vibrio cholerae when making the transition from an aquatic lifestyle to one more suited to a human host. An inverse correlation between motility and virulence gene expression has been reported, with the NADH : ubiquinone oxidoreductase system which powers motility by generat...
journal_title:Current opinion in microbiology
pub_type: 杂志文章,评审
doi:10.1016/s1369-5274(03)00032-8
更新日期:2003-04-01 00:00:00
abstract::Antibiotics started to be used almost 90 years ago to eradicate life-threatening infections. The urgency of the problem required rapid, broad-spectrum elimination of infectious agents. Since their initial discovery, these antimicrobials have saved millions of lives. However, they are not exempt from side effects, whic...
journal_title:Current opinion in microbiology
pub_type: 杂志文章,评审
doi:10.1016/j.mib.2017.05.014
更新日期:2017-06-01 00:00:00
abstract::Sexual development in filamentous ascomycetes requires mating-type genes to mediate recognition of compatible cell and nuclear types. Characterization of mating-type genes from various fungi shows that they primarily encode transcriptional regulators. Recent studies on mating-type-specific pheromones and internuclear ...
journal_title:Current opinion in microbiology
pub_type: 杂志文章,评审
doi:10.1016/s1369-5274(00)00073-4
更新日期:2000-04-01 00:00:00
abstract::Dynamic properties of type IV pili are essential for their function in bacterial infection, twitching motility and gene transfer. Laser tweezers are versatile tools to study the molecular mechanism underlying pilus dynamics at the single molecule level. Recently, these optical tweezers have been used to monitor pilus ...
journal_title:Current opinion in microbiology
pub_type: 杂志文章,评审
doi:10.1016/j.mib.2005.04.002
更新日期:2005-06-01 00:00:00
abstract::Organisms do not exist isolated from each other, but constantly interact. Cells can sense the presence of interaction partners by a range of receptors and, via complex regulatory networks, specifically react by changing the expression of many of their genes. Technological advances in next-generation sequencing over th...
journal_title:Current opinion in microbiology
pub_type: 杂志文章,评审
doi:10.1016/j.mib.2017.09.001
更新日期:2018-04-01 00:00:00
abstract::Most studies of small regulatory RNAs in bacteria have focussed on conserved transcripts in intergenic regions. However, several recent developments including single-nucleotide resolution transcriptome profiling by RNA-seq and increased knowledge of the cellular targets of the RNA chaperone Hfq suggest that the bacter...
journal_title:Current opinion in microbiology
pub_type: 杂志文章,评审
doi:10.1016/j.mib.2015.01.013
更新日期:2015-04-01 00:00:00
abstract::Recent work has shown that Immunoglobulin-like (Ig-like) domains occur frequently on the surface of tailed dsDNA bacteriophages. Several of these Ig-like domains are added to bacteriophage structural proteins via programmed ribosomal frameshifts, and their evolutionary patterns suggest that they can be exchanged by ho...
journal_title:Current opinion in microbiology
pub_type: 杂志文章,评审
doi:10.1016/j.mib.2007.05.018
更新日期:2007-08-01 00:00:00
abstract::The glideosome is a unique attribute of the Apicomplexa phylum. This myosin-based machine powers parasite motility, migration across biological barriers, host cell invasion and egress from infected cells. The timing, duration and orientation of gliding motility are tightly regulated to assure establishment of infectio...
journal_title:Current opinion in microbiology
pub_type: 杂志文章,评审
doi:10.1016/j.mib.2009.06.008
更新日期:2009-08-01 00:00:00
abstract::Mosquitoes are the major arthropod vectors of human diseases such as malaria and viral encephalitis. However, each mosquito species does not transmit every pathogen, owing to reasons that include specific evolutionary histories, mosquito immune system structure, and ecology. Even a competent vector species for a patho...
journal_title:Current opinion in microbiology
pub_type: 杂志文章,评审
doi:10.1016/j.mib.2012.04.001
更新日期:2012-06-01 00:00:00