Abstract：
:Targeted mutagenesis is one of the major tools for determining the function of a given gene and its involvement in bacterial pathogenesis. In mycobacteria, gene deletion is often accomplished by using allelic exchange techniques that commonly utilise a suicide delivery vector. We have adapted a widely-used suicide delivery vector (p1NIL) for cloning two flanking regions of a gene using ligation independent cloning (LIC). The pNILRB plasmid series produced allow a faster, more efficient and less laborious cloning procedure. In this paper we describe the making of pNILRB5, a modified version of p1NIL that contains two pairs of LIC sites flanking either a sacB or a lacZ gene. We demonstrate the success of this technique by generating 3 mycobacterial mutant strains. These vectors will contribute to more high-throughput methods of mutagenesis.
journal_name
J Microbiol Methodsjournal_title
Journal of microbiological methodsauthors
Balhana R,Stoker NG,Sikder MH,Chauviac FX,Kendall SLdoi
10.1016/j.mimet.2010.07.014subject
Has Abstractpub_date
2010-10-01 00:00:00pages
34-41issue
1eissn
0167-7012issn
1872-8359pii
S0167-7012(10)00245-9journal_volume
83pub_type
杂志文章abstract：UNLABELLED:An important goal for the improved diagnosis and management of infectious and inflammatory diseases, such as periodontitis, is the development of rapid and accurate technologies for the decentralized detection of bacterial pathogens. The aim of this prospective multicenter study was to evaluate the clinical ...
journal_title：Journal of microbiological methods
pub_type： 杂志文章,多中心研究
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abstract：:Yeast cell wall fractions have been proposed to bind enteropathogenic bacteria. The aim of this study was to develop a quantitative assay by measuring the optical density as growth parameter of adhering bacteria. The exponential growth phase of adhering bacteria was determined by optical density reading and compared w...
journal_title：Journal of microbiological methods
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abstract：:Systems that automate extraction of nucleic acid from cells or viruses in complex clinical matrices have tremendous value even in the absence of an integrated downstream detector. We describe our bench-top automated workstation that integrates our previously-reported extraction method - TruTip - with our newly-develop...
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abstract：:RNA-based molecular technique (RT-qPCR) is a promising method for microcystin monitoring in lakes and reservoirs, but great lability of RNA in cyanobacterial samples limits its application. To date, no studies have investigated how to effectively preserve RNA in cyanobacterial samples. In this study, four different tr...
journal_title：Journal of microbiological methods
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abstract：:A common method for visualizing bacterial biofilms is through confocal laser scanning microscopy images. Current software packages separate connected-biofilm bacteria from unconnected bacteria, such as planktonic or dispersed bacteria, but do not save both image sequences, making interpretation of the two bacterial po...
journal_title：Journal of microbiological methods
pub_type： 杂志文章
doi：10.1016/j.mimet.2012.04.008
更新日期：2012-07-01 00:00:00
abstract：:We have developed a method to identify and amplify differential fragments between two complex genomes. This technique, named hybridization-monitored genome differential analysis (HMDA), incorporates a monitor system into a PCR-based solid subtraction hybridization that tracks the entire hybridization process. This is ...
journal_title：Journal of microbiological methods
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abstract：:Whole-cell bacterial biosensors can be applied for the screening of antibacterial properties of extracts. We constructed a biosensor panel consisting of four different bacterial biosensor strains: Escherichia, Staphylococcus, Acinetobacter and Pseudomonas for expanded screening potential. The functionality of the pane...
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journal_title：Journal of microbiological methods
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journal_title：Journal of microbiological methods
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abstract：:Chemotactic behaviour is an important part of the lifestyle of motile bacteria and enables cells to respond to various environmental stimuli. The Hard Agar Plug (HAP) method is used to study the chemotactic behaviour of bacteria, including the fastidious microaerophile Campylobacter jejuni, an intestinal pathogen of h...
journal_title：Journal of microbiological methods
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doi：10.1016/j.mimet.2018.02.008
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abstract：:A sensitive, selective, and rapid protocol for detecting Ralstonia solanacearum from soil and plant tissues was developed based on the integration of the rapid self-replicating ability of bacteriophages with quantitative PCR (q-PCR). Six bacteriophages were isolated and selected for their ability to specifically infec...
journal_title：Journal of microbiological methods
pub_type： 杂志文章
doi：10.1016/j.mimet.2008.11.008
更新日期：2009-03-01 00:00:00
abstract：:The limited availability of biological samples hinders phylogenetic efforts to define structural differences among various biological groups. A novel workflow enabling the analysis of protists in low cell numbers by electron microscopy (EM) is described with cysts of Giardia intestinalis, a single-celled eukaryotic pa...
journal_title：Journal of microbiological methods
pub_type： 杂志文章
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abstract：:The susceptibility protocol established in this study takes into account that complex media are capable to buffer H2O2 which otherwise may adulterate test results. Furthermore, we demonstrate that clinical isolates of Staphylococcus aureus showed a higher resistance against H2O2 than Streptococcus pyogenes after 30 mi...
journal_title：Journal of microbiological methods
pub_type： 杂志文章
doi：10.1016/j.mimet.2018.11.001
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abstract：:The quality and quantity of sputum collected has an important impact on the laboratory diagnosis of pulmonary TB. We conducted a pilot study to assess a new collection cups for the collection of sputum for the diagnosis of pulmonary tuberculosis. The pilot study utilized the standard collection cup in South Africa dem...
journal_title：Journal of microbiological methods
pub_type： 杂志文章
doi：10.1016/j.mimet.2017.01.011
更新日期：2017-04-01 00:00:00
abstract：:Cyanobacteria are an ancient and diverse group of photosynthetic microorganisms, which inhabit many different and extreme environments. This indicates a high degree of biological adaptation, which has enabled these organisms to thrive and compete effectively in nature. The filamentous cyanobacterium, Lyngbya majuscula...
journal_title：Journal of microbiological methods
pub_type： 杂志文章
doi：10.1016/s0167-7012(01)00324-4
更新日期：2002-02-01 00:00:00
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journal_title：Journal of microbiological methods
pub_type： 杂志文章
doi：10.1016/j.mimet.2010.03.026
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abstract：:The cDNA conversion of RNA molecules is a prerequisite for their analysis. In the case of prokaryotic RNAs, cDNA conversion is difficult due to a lack of the long poly(A) tails that are found in eukaryotic mRNAs. The full cDNAs for eukaryotic mRNAs can be amplified by the reverse transcription polymerase chain reactio...
journal_title：Journal of microbiological methods
pub_type： 杂志文章
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abstract：:In wastewater, biological phosphate removal can fail because of the presence of glycogen accumulating organism (GAO), therefore measuring glycogen stored in microbial cultures provides information on the bacterial population type. Once glycogen is hydrolysed to glucose it was accurately measured using a human glucose ...
journal_title：Journal of microbiological methods
pub_type： 杂志文章
doi：10.1016/j.mimet.2014.03.002
更新日期：2014-05-01 00:00:00
abstract：:Burkholderia pseudomallei is the causative agent of melioidosis, a potentially fatal disease endemic or emerging world-wide. Here we report unmarked allele-replacement mutagenesis using efficient sacB counter-selection. Despite being genotypically sacB(+), most commonly used B. pseudomallei strains are sucrose-resista...
journal_title：Journal of microbiological methods
pub_type： 杂志文章
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abstract：:The S. PneumoStrip test is a recently developed reverse hybridization strip-based commercial assay that allows for the identification of 76 pneumococcal serotypes, 42 individually and 34 in pairs, according to their specific gene sequences. The test was validated with reference strains of 92 different pneumococcal ser...
journal_title：Journal of microbiological methods
pub_type： 杂志文章
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journal_title：Journal of microbiological methods
pub_type： 杂志文章
doi：10.1016/j.mimet.2018.09.007
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abstract：:Loop-mediated isothermal amplification (LAMP) is considered as one of the alternatives to the conventional PCR and it is an inexpensive portable diagnostic system with minimal power consumption. The present work describes the application of LAMP in real-time photon detection and quantitative analysis of nucleic acids ...
journal_title：Journal of microbiological methods
pub_type： 杂志文章
doi：10.1016/j.mimet.2016.12.002
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abstract：:The analysis of soil fungal communities by molecular fingerprinting and subsequent identification of the underlying populations require the amplification of a phylogenetically informative gene fragment. In this study we tested the reliability and suitability of the previously published fungal primer combination (NS1/F...
journal_title：Journal of microbiological methods
pub_type： 杂志文章
doi：10.1016/j.mimet.2005.06.014
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abstract：:Anthrax is one of the important diseases in humans and animals, caused by the gram-positive bacteria spores called Bacillus anthracis. The disease is still one of the health problems of developing countries. Due to fatigue and decreased visual acuity, microscopic diagnosis of diseases by humans may not be of good qual...
journal_title：Journal of microbiological methods
pub_type： 杂志文章
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abstract：:Identifying microorganisms that are active under specific conditions in ecosystems is a challenge in microbial ecology. Recently, the bromodeoxyuridine (BrdU) technique was developed to label actively growing cells. BrdU, a thymidine analog, is incorporated into newly synthesized DNA, and the BrdU-labeled DNA is then ...
journal_title：Journal of microbiological methods
pub_type： 杂志文章
doi：10.1016/j.mimet.2010.09.016
更新日期：2010-12-01 00:00:00
abstract：:Vibrio parahaemolyticus is one of the important foodborne pathogens is of public health concern due to the emergence of pandemic strains causing disease outbreaks worldwide. We evaluated the DNA based colony hybridization technique for the detection and enumeration of total and pathogenic V. parahaemolyticus from the ...
journal_title：Journal of microbiological methods
pub_type： 杂志文章
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abstract：:MTS or {3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl}-2H-tetrazolium, inner salt) is converted into soluble formazan by mitochondrial dehydrogenase of viable cells, thus serving as an indicator of cell viability. Accordingly, a MTS-based assay was developed to evaluate anti-leishmanial acti...
journal_title：Journal of microbiological methods
pub_type： 杂志文章
doi：10.1016/j.mimet.2005.10.011
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abstract：:The lack of a surrogate-of-immunity assay presents a major barrier to Streptococcus pyogenes research. Modification of the Lancefield assay to include an antibody digestion step reduced inter-donor variation and permitted detection of the anti-streptococcal activity of intravenous immunoglobulin and convalescent serum...
journal_title：Journal of microbiological methods
pub_type： 杂志文章
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abstract：:Recently, a unique method for bacterial transformation using nanofibers to inoculate DNA has been developed by Naoto Yoshida and colleagues. We have verified the principle, transforming Escherichia coli, Yersinia enterocolitica and Acinetobacter baumannii, and have established a user-friendly protocol. A buffered susp...
journal_title：Journal of microbiological methods
pub_type： 杂志文章
doi：10.1016/j.mimet.2009.12.002
更新日期：2010-02-01 00:00:00
