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Transformation, Bacterial (0):
Recent article
Transformation, Bacterial
Biomacromolecules (12)9 2011
Abstract
Polyhydroxyalkanoates (PHA) synthesis genes phbC and orfZ cloned from Ralstonia eutropha H16 were transformed into beta-oxidation weakened Pseudomonas putida KTOY08ΔGC, a mutant of P. putida KT2442. The recombinant P. putida strain termed KTHH06 was able to produce a short-chain-len...
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PMID: 21863836
PDF is available here.
PDF is available here.
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Abstract
We cloned genes coding for human HMGB1 and its active components A box and B box by PCR and inserted the cloned genes into pET28a vectors for transformation of Escherichia coli BL21. The E. coli expressed proteins were then purified with a Ni(2+)-NTA column and the endotoxin content was removed. Rec...
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PMID: 21255697
PDF is available here.
PDF is available here.
Abstract
To prepare and purify recombinant human NAMPT and NAMPT (H247A) proteins and to detect their enzymatic activity.
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PMID: 21488211
PDF is available here.
PDF is available here.
Abstract
Fatty-acid binding proteins (FABPs) are abundantly expressed proteins that bind a range of lipophilic molecules. They have been implicated in the import and intracellular distribution of their ligands and have been linked with metabolic and inflammatory responses in the cells in which they are expre...
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PMID: 21301109
PDF is available here.
PDF is available here.
Abstract
The cysteine protease inhibitor from Ascaris lumbricoides, a roundworm that lives in the human intestine, may be involved in the suppression of human immune responses. Here, the molecular cloning, protein expression and purification, preliminary crystallization and crystallographic characterization...
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PMID: 21301092
PDF is available here.
PDF is available here.
Abstract
Phospholipases D are the major dermonecrotic component of Loxosceles venom and catalyze the hydrolysis of phospholipids, resulting in the formation of lipid mediators such as ceramide-1-phosphate and lysophosphatidic acid which can induce pathological and biological responses. Phospholipases D can b...
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PMID: 21301094
PDF is available here.
PDF is available here.
Abstract
The Toll/interleukin-1 receptor (TIR) domain is a protein-protein interaction domain that is found in both animal and plant immune receptors. In animal Toll-like receptor signalling, both homotypic TIR-domain interactions between two receptor molecules and heterotypic interactions between receptors...
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PMID: 21301095
PDF is available here.
PDF is available here.
Abstract
We report here a versatile gene expression method for the genetic engineering of R. eutropha. This method, based on a simplified electroporation protocol, uses a recombinant plasmid, pBS29-P2, containing a Pseudomonas syringae promoter (P2) and two antibiotic-resistance markers (i.e., genes coding f...
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PMID: 20447426
PDF is available here.
PDF is available here.
Abstract
We report the design, synthesis, and assembly of the 1.08-mega-base pair Mycoplasma mycoides JCVI-syn1.0 genome starting from digitized genome sequence information and its transplantation into a M. capricolum recipient cell to create new M. mycoides cells that are controlled only by the synthetic ch...
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PMID: 20488990
PDF is available here.
PDF is available here.
Infection and Immunity (78)7 2010
Abstract
We have recently started a systematic functional analysis of the genes involved in Tfp biology in a well-characterized clinical isolate of the human pathogen Neisseria meningitidis. After previously studying 16 genes involved in Tfp biogenesis, here we report the characterization of 7 genes that are...
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PMID: 20439474
PDF is available here.
PDF is available here.
Abstract
We report evidence of natural genetic competence in 13 Thermoanaerobacter and Thermoanaerobacterium strains previously believed to be difficult to transform or genetically recalcitrant. In Thermoanaerobacterium saccharolyticum JW/SL-YS485, natural competence-mediated DNA incorporation occurs during...
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PMID: 20472726
PDF is available here.
PDF is available here.
Current Microbiology (61)1 2010
Abstract
The marine bacterium Neptuniibacter sp. strain CAR-SF utilizes carbazole as its sole carbon and nitrogen sources. Two sets of clustered genes related to carbazole degradation, the upper and lower pathways, were obtained. The marine bacterium genes responsible for the upper carbazole degradation path...
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PMID: 20039169
PDF is available here.
PDF is available here.
Current Microbiology (61)1 2010
Abstract
We generated three auxotrophic recombinant L. lactis subsp. lactis IL1403 strains expressing a heterologous protein, BmpB, using thyA- and alr-targeting integration vectors: ITD (thyA(-)alr(+) bmpB(+)), IAD (thyA(+)alr(-)bmpB(+)), and ITDAD (thyA(-)alr(-) bmpB(+)). After construction of integration...
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PMID: 20035330
PDF is available here.
PDF is available here.
The Lancet (375)9730 2010
Abstract
Abstract
The different cleavage patterns of pYBamy59 plasmid isolated from E. coli DH5alpha and B. longum MG1 by the cell extract of B. longum MG1 suggested that the main reason for low transformation efficiency was related to the restriction modification (R-M) system. To confirm correlation between R-M syst...
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PMID: 20622503
PDF is available here.
PDF is available here.
Abstract
Abstract
To clone and produce ribosome inactivating protein MAP30 from the seeds of Momordica charantia L(bitter melon), and to evaluate the biological activity of the recombinant protein.
The DNA sequence encoding MAP30 was cloned from the fresh seeds of Momordica charantia by PCR, the target DNA fragments...
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PMID: 20544988
PDF is available here.
PDF is available here.
Abstract
To construct COL1A1-targeted short hairpin RNA (shRNA) vector with pSilencer 4.1-CMV neo siRNA expression vector and to evaluate its effect on proliferation and migration of gastric cancer BGC-823 cells in vitro.
Three COL1A1-shRNA plasmids (COL1A1-shRNA-1, COL1A1-shRNA-2, COL1A1-shRNA-3), targeting...
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PMID: 20544987
PDF is available here.
PDF is available here.
Journal of Bacteriology (192)4 2010
Abstract
We previously showed that a periplasmic DNase, encoded by dns, inhibits natural transformation in C. jejuni. In the present study, genetic factors coding for DNase activity in the absence of dns were identified. DNA arrays indicated that nonnaturally transformable dns-negative strains contain putati...
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PMID: 20023031
PDF is available here.
PDF is available here.
Euro surveillance : bulletin europeen sur les maladies transmissibles = European communicable disease bulletin (15)1 2010
Abstract
We document here the in vivo transfer of bla(KPC-2) between intensive care unit-acquired and a commensal strain of K. pneumoniae in a French patient after his repatriation from Greece. This first report of an emerging KPC-producing strain in France raises further concerns about the spread of carbape...
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PMID: 20067746
PDF is available here.
PDF is available here.
Abstract
The CaCl2 method, commonly used for transformation of Escherichia coli, was modified and used to develop a simpler and easier transformation method for Rhizobia sp. Two species of Rhizobia, Sinorhizobium meliloti MTCC 100 and Mesorhizobium ciceri TAL 620, were transformed with the 13.2 kb binary vec...
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PMID: 20130696
PDF is available here.
PDF is available here.
Abstract
We provide the experimental single crystal X-ray diffraction results together with theoretical methods to characterize the hydrolysis/decomposition reactions of taurolidine. The crystal structure features two independent molecules linked through intermolecular H-bonds with one of them somewhat posit...
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PMID: 20126631
PDF is available here.
PDF is available here.
Abstract
We present protocols for natural transformation in strains found in the three groups of transformable oral streptococci, with focus on methods using synthetic CSPs. We also include suggestions on how to optimize competence conditions for individual species or strains....
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PMID: 20717785
PDF is available here.
PDF is available here.
Abstract
Acidithiobacillus caldus is an acidophilic, chemoautotrophic bacterium, plays important role in bioleaching. Gene transformation into A. caldus is difficult and only the conjugation method was reported successful, which method was relatively sophisticated. In this research, electrotransformation of...
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PMID: 20134231
PDF is available here.
PDF is available here.
Abstract
We had previously determined by substrate binding test in vitro. The result of this study indicates that type I PKS-SAT can alter its specificity in vivo and functions well in extender units and proved the feasibility of the SAT-EAT domain replacement in type I PKS. We propose that SAT-EAT replaceme...
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PMID: 20391758
PDF is available here.
PDF is available here.
Abstract
Glutathione S-transferases (GSTs) are a family of multifunctional enzymes involved in cellular detoxification that catalyze the attachment of electrophilic substrates to glutathione. Two classes of GSTs related to the rho and sigma classes of enzymes in Antarctic bivalves have been c...
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PMID: 19651242
PDF is available here.
PDF is available here.
Methods in Molecular Biology (576)576 2009
Abstract
We describe a manual microdissection strategy for the isolation of high-quality RNA. Furthermore, a strategy for combining linear amplification of RNA with longSAGE is described that allows the use of antisense RNA (aRNA) generated via the well-established linear amplification of RNA procedure toget...
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PMID: 19882261
PDF is available here.
PDF is available here.
Abstract
The efficiencies of different transformation methods of E. coli DH5Qalpha train, induced by several cations like Mg2+, Mn2+ Rb+ and especially Ca2+, with or without polyethylene glycol (PEG) and dimethyl sulfoxide (DMSO) were compared using the two commonly used plasmid vectors pCAMBIA1201 and pBI12...
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PMID: 20027870
PDF is available here.
PDF is available here.
Analytical Biochemistry (394)2 2009
Abstract
We report the successful development of a coupled, continuous spectrophotometric assay to measure the activity of ChlM. The product of the methyltransferase reaction, S-adenosyl-l-homocysteine (SAH), is converted into adenine and then hypoxanthine by the recombinant coupling enzymes SAH nucleosidase...
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PMID: 19646414
PDF is available here.
PDF is available here.
Abstract
To study the anti-platelet GPVI single chain Fv phage antibody which can inhibit the aggregation function of platelet by using phage antibody library technology.
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PMID: 19954615
PDF is available here.
PDF is available here.
Analytical Biochemistry (393)2 2009
Abstract
Our goal was to design automated protocols that would allow the purification of proteins with minimal operator intervention. Separate methods have been produced and tested that automate the sample loading, column washing, sample elution and peak collection steps for ion exchange, metal affinity, hyd...
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PMID: 19595984
PDF is available here.
PDF is available here.
Analytical Biochemistry (394)2 2009
Abstract
We present a new method for tracking translation steps in cell-free protein synthesis using atomic force microscopy (AFM). The cell-free protein synthesis system is useful to track the inherent translational progress of a target gene, whereas conventional UV absorption measurement coupled with densi...
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PMID: 19643072
PDF is available here.
PDF is available here.
Abstract
The cloned gene fragment exhibited complete similarity with the published sequence of gra8 gene by sequence analysis. rGRA8 was expressed in Escherichia coli in fusion with a very small tag and the soluble protein was purified by immobilized metal ion affinity chromatography. In immunoblot, serum sa...
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PMID: 19688020
PDF is available here.
PDF is available here.
Abstract
We investigated the efficiency of various DNA types including chromosomal linear DNA and circular or linearized integrative and (or) replicative plasmids to naturally transform R. solanacearum. To study the respective regulatory role of DNA transport and maintenance in the definite acquisition of ne...
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PMID: 19767847
PDF is available here.
PDF is available here.
Antonie Van Leeuwenhoek (96)1 2009
Abstract
Bacillus thuringiensis HD-73 was transformed with the endochitinase gene chiA74 under the control of a strong promoter (pcytA) and a 5' mRNA stabilizing (STAB-SD) sequence (HD-73-pEBchiA74). Expression levels were compared with those observed from the wild type strain (HD-73) and the recombinant HD-...
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PMID: 19337851
PDF is available here.
PDF is available here.
Abstract
In 2006, a new variant of Chlamydia trachomatis (nvCT) was reported in Sweden. The nvCT contains a deletion that includes the targets for the C. trachomatis genetic diagnostic single-target systems from Roche Diagnostics and Abbott Laboratories. Roche and Abbott have now developed certified dual-tar...
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PMID: 19442398
PDF is available here.
PDF is available here.
Abstract
The DNA encoding keratinase from Pseudomonas aeruginosa was ligated into pET-43b(+) expression vector and transformed into Escherichia coli AD494(DE3)pLysS. After isopropyl beta-d-thiogalactopyranoside induction, the soluble recombinant keratinase was expressed in E. coli. The keratinase with a mole...
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PMID: 19415925
PDF is available here.
PDF is available here.
Abstract
We therefore suggest that the Mob function in pLFE1 is supplied in trans from another plasmid present in L. plantarum M345.Filter-mating experiments showed that pLFE1 has a broad host-range with transconjugants obtained from Lactobacillus rhamnosus, Lactococcus lactis, Listeria innocua, the opportun...
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PMID: 19399991
PDF is available here.
PDF is available here.
Abstract
We conclude that the first restriction endonuclease, designated BbrI, is an isoschizomer of BbeI, the second, BbrII, is a neoschizomer of SalI, while the third, BbrIII, is an isoschizomer of PstI. Expression of each of the B. breve UCC2003 methylase-encoding genes in B. breve JCM 7017 established th...
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PMID: 21261927
PDF is available here.
PDF is available here.
Microbiology (155)Pt 4 2009
Abstract
Deletions leading to complete or partial removal of ParB were introduced into the Pseudomonas aeruginosa chromosome. Fluorescence microscopy of fixed cells showed that ParB mutants lacking the C-terminal domain or HTH motif formed multiple, less intense foci scattered irregularly, in contrast to the...
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PMID: 19332810
PDF is available here.
PDF is available here.
Current Microbiology (58)4 2009
Abstract
In the development of an oral vaccine against Helicobacter pylori, H. pylori urease subunit B (UreB) was expressed in a food-grade delivery vehicle, Lactococcus lactis NZ3900. The ureB gene (Genbank accession no. FJ436980) was amplified by polymerase chain reaction (PCR) from MEL-Hp27. The PCR-ampli...
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PMID: 19238485
PDF is available here.
PDF is available here.
Abstract
A new cloning system is described, which allows the construction of large-insert fosmid libraries in Escherichia coli and the transfer of the recombinant libraries to the extreme thermophile Thermus thermophilus via natural transformation. Libraries are established in the thermophili...
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PMID: 19285378
PDF is available here.
PDF is available here.
Microbiology (155)Pt 3 2009
Abstract
We searched for DNA binding proteins within the meningococcal inner membrane. Inner membrane fractions from a panel of neisserial strains were subjected to a solid-phase overlay assay with DNA substrates, and MS was subsequently employed to identify proteins that bind DNA. A number of DNA binding co...
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PMID: 19246756
PDF is available here.
PDF is available here.
A novel dichloromethane-degrading Lysinibacillus sphaericus strain wh22 and its degradative plasmid.
Abstract
Dichloromethane (DCM)-degrading bacterium strain wh22 (GenBank accession number FJ418643) was isolated and identified as Lysinibacillus sphaericus based on standard morphological and physiological properties, cellular fatty acid composition, mole percent guanine-cytosine content, and nucleotide sequ...
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PMID: 19183982
PDF is available here.
PDF is available here.
Abstract
We constructed an Escherichia coli to B. thuringiensis shuttle vector, pEMB0557, with a large loading capacity. This vector incorporated the ori60 replicon from B. thuringiensis subsp. kurstaki YBT-1520, erythromycin resistance (B. thuringiensis), and chloromycetin resistance (E. coli) genes. A bact...
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PMID: 19153728
PDF is available here.
PDF is available here.
Abstract
These results indicate that Agrobacterium functional screening is an effective supplemental method of pre-screening genes involved in abiotic stress tolerance....
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PMID: 19202291
PDF is available here.
PDF is available here.
Abstract
Although phosphatidylserine synthase (PSS) from Escherichia coli is an ideal enzyme for phospholipid production, its application in the food industry has been limited because of the low PSS yield. In this study, the pss gene was cloned from E. coli K(12) and expressed in Bacillus sub...
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PMID: 19072541
PDF is available here.
PDF is available here.
Abstract
We demonstrate an easy and cost effective way to introduce base substitutions into a plasmid. The protocol works with standard reagents and is independent from commercial kits, which often are very expensive. Applying this protocol can reduce the total cost of a reaction to an eighth of what it cost...
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PMID: 19488024
PDF is available here.
PDF is available here.
Abstract
A method is described for the identification of the antigen recognised by an scFv isolated from an antibody phage display library using selection against a complex mixture of proteins (e.g. intact cells, purified cell surface membranes, and tissue sections). The method takes advantage of a yeast two...
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PMID: 19554295
PDF is available here.
PDF is available here.
Abstract
We previously described the establishment of a binary vector system that allows co-expression of SUMO conjugation enzymes and a target protein of interest, leading to efficient SUMO modification and the production of a large amount of recombinant SUMO-modified proteins in Escherichia coli. The advan...
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PMID: 19107420
PDF is available here.
PDF is available here.
Abstract
RNA interference is a phenomenon in which specific, endogenous genes are silenced by mRNA degradation. This technology is highly regarded as a potential therapeutic due to its high efficacy and low toxicity. However, the difficulty of delivering RNAi to target cells has impeded the development of RN...
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PMID: 19048211
PDF is available here.
PDF is available here.
Abstract
We describe methods that have been optimized for the FYVE and phox (PX) domains of the EEA1 and Vam7p proteins, respectively, both of which specifically recognize phosphatidylinositol 3-phosphate (PtdIns3P) within endocytic membranes. Solution-state NMR methods were used to characterize the phosphoi...
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PMID: 19160678
PDF is available here.
PDF is available here.
Microbiology (155)Pt 1 2009
Abstract
We describe how purified SufA efficiently and specifically cleaves fibrinogen in human plasma. SufA is both secreted by F. magna and associated with the bacterial surface. Successful gene targeting has previously not been performed in anaerobic cocci, but in order to study the role of the SufA that...
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PMID: 19118364
PDF is available here.
PDF is available here.
Abstract
We amplified this gene from the H37Rv genome, and His-tagged Rv0901 protein was expressed and purified. Also, a recombinant plasmid bearing Rv0901 was constructed and electroporated into a virulent Mycobacterium smegmatis, using shuttle expression vector pMV261. Transformants were induced to express...
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PMID: 19168955
PDF is available here.
PDF is available here.
Abstract
We report the nucleotide sequence of two novel cryptic plasmids (4357 and 14 662 base pairs) carried by a Yersinia enterocolitica biotype 1A strain isolated from pork. As distinguished from most biotype 1A strains, this isolate, designated 07-04449, exhibited adherence to eukaryotic cells. The small...
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PMID: 19834620
PDF is available here.
PDF is available here.
BMC Microbiology (9) 2009
Abstract
In presence of ampicillin (AMP), S. typh-lux with pCGLS-1, pAK1-lux and pXEN-1 plasmids exhibited 100% photon-emitting colonies over a 10-d study period. Photon emitters of S. typh-lux with pCGLS-1, pAK1-lux and pXEN-1 without AMP selection decreased over time (P < 0.05), representing only 11 +/- 1%...
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PMID: 19635131
PDF is available here.
PDF is available here.
Journal of Antibiotics (62)1 2009
Abstract
Resistance to metronidazole (MTZ) in Helicobacter pylori is associated with mutations in rdxA, encoding an oxygen-insensitive NADPH nitroreductase, and mutations in frxA, encoding a NAD(P)H-flavin oxidoreductase. Despite this association, the strict correlation of MTZ resistance with mutations in rd...
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PMID: 19132060
PDF is available here.
PDF is available here.
Abstract
We show that CRISPR interference prevents conjugation and plasmid transformation in S. epidermidis. Insertion of a self-splicing intron into nickase blocks interference despite the reconstitution of the target sequence in the spliced mRNA, which indicates that the interference machinery targets DNA...
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PMID: 19095942
PDF is available here.
PDF is available here.