How To Use Plasmid In A Sentence
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Single radiolabelled colonies harbouring plasmids with inserts bigger than 1000 bp were analysed further.
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Methods: The restriction endonuclease and T 4 DNA ligase were used to construct the vector plasmid.
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Results PCR, restriction endonuclease method and direct sequence analysis demonstrated that plasmids of YMDD, YVDD and YIDD were constructed successfully.
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Plasmid DNA was prepared using the Qiagen plasmid purification system.
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Agrobacteria gene movements have produced second chromosomes derived from plasmids, while in the biovar II strain K84 the plasmid-based replicon has yet to reach second chromosome status.
Health News from Medical News Today
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Yersinia pestis in the 109 samples from the cemetery in East Smithfield, his lab employed a sort of sensitive fishing technique, using tiny segments of DNA that matched up with segments from a ring of DNA, called a plasmid, found in the bacterium.
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1987 (Burke et al. 1987) permitted much larger segments of DNA to be ordered and stored for sequencing than was possible with plasmid or cosmid libraries.
The Human Genome Project
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Objective To construct the plasmid expressing BDV p 40 gene.
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Biological containment involves crippling the plasmid vector and its bacterial host so that they will not be able to survive outside the laboratory.
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That vehicle (often a subcellular structure called a plasmid) is then inserted into a different cell, which activates the gene, leading to the production of a scientifically or commercially useful protein.
Scientists create cell based on man-made genetic instructions
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Similarly, a Cre-expressing plasmid or adenovirus was employed to remove three or four copies of a polycistronic vector from the genome of mouse iPSCs
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Results The protein expressed by the plasmid which had a correct reading frame, had no transcriptional activation and could interact with a known coactivator PNRC.
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Conjugative plasmids can mediate gene transfer between bacterial taxa in diverse environments.
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This fragment was cloned into a plasmid vector and sequenced to exclude mutations introduced during PCR.
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Any genetic vectors suitable for transformation of the cells of interest may be employed, including but not limited to adenoviral vectors, plasmids, and non-viral vectors, such as cationic lipids.
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The plasmid must contain prokaryotic nucleotide sequences coding for a bacterial replication origin for DNA and an antibiotic resistance gene.
Archive 2005-10-01
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Genes of interest were printed in duplicate along with non-specific genes (plasmid c-DNA) and housekeeping genes were used for normalization purposes.
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The fibroblast line grows adherently to glass and plastic in culture and is generally utilized as a transfection host for virus genomes and recombinant plasmids.
Undefined
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The value for the ancestor was expressed relative to the midpoint topoisomer of plasmid that was completely relaxed by using calf-thymus topoisomerase I and assigned = 0.
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Escherichia coli (strain EN222 which contains the structural gene for the enzyme on a multicopy plasmid [1]; strain GR143 and EN222, and strain which produce recombinant enzymes
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Schneider cells were seeded onto cover slips and after 24 hr the cells were transfected with plasmid pBSHS-Fl d.
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Objective Construct a recombinant plasmid pET 28 a - EDA - EDB , prepare the fusion EDA - EDB protein.
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The researchers injected the DNA plasmid, which encodes a gene that stimulates the immune system, directly into the tumor site in mice.
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While E. coli carries its main genetic archive on conventional chromosomes, plasmids equip the microbe with small libraries of supplementary information.
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Methods Isolates were identified as acinetobacter calcoaceticus by using antibiotic susceptibility test, plasmid profiles, restriction enzyme fingerprinting assay and plasmid elimination method.
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Other analyses have inspected the interplay between similar selective mechanisms involved in the reproduction of viruses, prebiotic replicators, and cytoplasmic organelles, but do not deal with plasmid-specific traits.
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PC - 3 cells were transfected with mGM - CSF expressing plasmid or the control vector by lipo - some transfection.
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He has played the neological game himself with entries like plasmid, exobiology, euphenics, and prototroph.
The Scientist
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The predicted 5' and 3' boundaries of the expression regions were confirmed by direct plasmid DNA sequencing.
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The proportion of recombinant plasmids in each sample was then estimated by comparing the two numbers.
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The screening of human tissue plasminogen activator recombinant plasmid showed that the two methods can recognize the positive colones quickly and correctly.
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The incorporation of plasmids or prophages into the bacterial chromosomes is called transfection.
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Indeed, atypical E. coli that grow aerobically on citrate (Cit+) have been isolated from agricultural and clinical settings, and were found to harbor plasmids, presumably acquired from other species, that encode citrate transporters (44, 45).
Assessing Fault
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The lactic acid bacteria naturally contain from one to more than ten distinct plasmids, and metabolically important traits, including lactose-fermenting ability, bacteriophage resistance, and bacteriocin production, have been linked to plasmid DNA.
1 Upgrading Traditional Biotechnological Processes
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This conclusion was further supported by the fact that large part of the secretome was found to be coded by a piece of extra-chromosomal DNA called plasmid, which is, simply, the most mobile part of the whole bacterial genome
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The gene transference involves the use of a vector carrier which can be a plasmid or a virus.
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Liposomal formulations of genetic drugs such as antisense oligonucleotides and plasmid DNA are difficult to achieve.
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An increase was seen in all 7 rabbits transfected with alkaline phosphatase plasmid.
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The nucleotide sequences of the constructed plasmids were verified by DNA sequencing.
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Unwanted materials, including RNA, DNA, and proteins, are thoroughly washed from the column before the plasmid DNA is eluted using a high salt buffer.
Archive 2005-10-01
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The recombinant plasmid PET- 11D-ospC was identified with restriction endonuclease analysis and sequencing.
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Objective Construct a recombinant plasmid pET 28 a - EDA - EDB , prepare the fusion EDA - EDB protein.
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Twenty-four hours after transfection, the cells were harvested and the plasmid DNA recovered.
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After extracting with phenol-chloroform, it is not necessary to recover the plasmid by ethanol precipitation.
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Alcohol (isopropanol) precipitation concentrates the eluted plasmid DNA, which is collected by centrifugation, washed, and redissolved in buffer.
Archive 2005-10-01
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C. Resistance to penicillin is mediated both by plasmid-encoded penicillinase and by changes in chromosome-encoded proteins that control the permeability of the drug.
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For each transformation, the plasmid inserts from 7 to 18 colonies were sequenced.
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Figure 2 . Restriction pattern of the recombinant plasmid.
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Co-transfection of human TLR2 with increasing amounts of the expression plasmid encoding alphaT2ib into HEK293 cells demonstrated high efficiency of the TLR2-alphaT2ib interaction.
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For many years, the method of choice for supercoiled plasmid and virus DNA purification was cesium chloride density gradient centrifugation in the presence of an intercalation agent (such as ethidium bromide or propidium iodide).
Archive 2005-10-01
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This plasmid is optimal for nonmammalian expression. pDsRed1-1: This plasmid contains an RFP gene that has been altered for human expression of RFP (DsRed1).
Archive 2005-10-01
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In this environment, I began the transition to experimental molecular biology by using the heteroduplex method and electron microscopy to study the structure of plasmids of the sex factors and drug resistant factors of bacteria.
Phillip A. Sharp - Autobiography
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The bacterium contains a plasmid, a circular piece of DNA that holds the gene and promoter sequence.
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However, the genes brought in by a plasmid do not usually become a permanent part of the bacterium's own chromosome.
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The replication control genes of bacterial plasmids face selection at two conflicting levels.
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Fertility genes enable a plasmid's genetic information to be transferred from a donor to a recipient strain.
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Recombinant plasmids were purified from overnight cultures using the alkaline lysis technique.
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The only apparent regulatory difference is a short time delay when activators reside in the cytoplasm before binding to plasmids.
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Plasmid manipulation and bacterial transformation were performed by standard techniques.
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For many years, the method of choice for supercoiled plasmid and virus DNA purification was cesium chloride density gradient centrifugation in the presence of an intercalation agent (such as ethidium bromide or propidium iodide).
Archive 2005-10-01
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The two ORFs downstream of hlyF share homology with the two aforementioned avian plasmids and a recently sequenced plasmid p1658 / 97 from a human septicemic hlyF has been identified in E. coli strains isolated from broilers
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The application to rats of recombinant human insulin gene retroviral vector plasmid mutated in three place with physiological regulatory element constructed by us was safe.
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The sorted cells were then subjected to limiting dilution to obtain pure clones of double-positive cells, followed by electroporation of a Flp-expression plasmid to achieve a clean translocation.
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Alcohol (isopropanol) precipitation concentrates the eluted plasmid DNA, which is collected by centrifugation, washed, and redissolved in buffer.
Archive 2005-10-01
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Recently, simplified ion-exchange column chromatography methods (commonly termed a mini-prep) have largely supplanted the cumbersome and time-consuming centrifugation protocol to yield large quantities of endotoxin-free plasmid DNA in a relatively short period of time.
Archive 2005-10-01
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Single radiolabelled colonies harbouring plasmids with inserts bigger than 1000 bp were analysed further.
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We hypothesized that the highly efficient plasmid-based piggyBac transposon system would enable long-term inducible gene expression in mice in vivo.
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It's a type of resistance that's carried on a mobile piece of DNA called a plasmid, and it's able very quickly to transfer from bacteria to bacteria," said Walsh.
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Each step in the curing procedures was monitored by electrophoresis of plasmid isolations.
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Genetic backcrosses were conducted to eliminate plasmid-based mutations and to verify that a single genetic locus was responsible for the 3 - AT-resistant phenotype.
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Spontaneous SOS gene expression was measured in strain GY7109 recA carrying plasmids with different recAX chimeras.
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Illustrated in Figure 7 is the restriction enzyme and genetic map of a commercially available (BD Biosciences Clontech) bacterial plasmid derivative containing the coding sequence for enhanced yellow fluorescent protein fused to the endoplasmic reticulum targeting sequence of calreticulin (a resident protein).
Archive 2005-10-01
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To select for the plasmids (donors or transconjugants), we used kanamycin and chloramphenicol.
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Molecular access to the sda gene was via the sda HZ.P1 mutation by the method of plasmid rescue of genomic DNA flanking the transposon insertion.
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Some are encoded in bacterial plasmids and mediate bacterial conjugation, or, in the case of Agrobacterium tumefaciens, T-DNA transport into plant cells.
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In particular, the position of CpG sites (upstream, around or downstream of TSS) deserves further attention since it had been shown that DNA methylation downstream of TSS can impact transcriptional elongation at integrated transgenes or episomal plasmids
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Four lysogens (HI2678, HI2679, HI2680, and HI2681) were prepared by transformation of the wild-type strain or ligts7 mutant strain with the high-copy-number plasmids pSY343 or pLP312.
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The genotype of each strain and plasmids used for transformation are indicated at the top.
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Positive phage clones were converted to plasmid vectors using the material provided in the cDNA synthesis kit.
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For plasmids, you'll have the standard assortment of abilities like electrobolt and incinerate, as well as something called aero dash which lets you surge forward at high speed and, if you make contact with a frozen target, break them to pieces.
IGN Complete
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Harvested bacterial colonies containing mutant plasmids were grown in LB medium in the presence of ampicillin and kanamycin to maintain the selective pressure on the mutants.
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Purified ATF - lys 10 specifically bound to uPAR - expressing cells and formed protein - DNA complexes with plasmid pGL 3 - control.
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A Tol2 clone marked with the bacterial tetracycline-resistance gene was microinjected into fertilized eggs together with a target plasmid, and the plasmid was recovered from embryos.
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Insert and ligate any DNA fragment into a vector such as plasmid or viral vector, to form a recombinant DNA.
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The solid line designates a plasmid that conjugates slowly but imposes a low cost of carriage, whereas the dashed line indicates a faster-conjugating plasmid that imposes a higher cost.
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By restriction enzyme identification, PCR and sequence analysis, the recombinant plasmid of the HA1 was successfully constructed.
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The accuracy of each mutagenized plasmid was checked by sequencing.
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Plasmid: circular DNA molecules able to replicate independently of the chromosome in microorganisms.
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We note, however, that few if any of the genes borne by bacterial plasmids are under strong positive selection at all times.
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The complete absence of untransformed animals in these transformed lines suggested that these stable lines were homozygous for a chromosomal insertion of the transforming plasmid.
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Methods: The restriction endonuclease and T 4 DNA ligase were used to construct the vector plasmid.
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Plasmid transformation, selection, and testing for recombinant clones were performed as described.
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According to recent studies, most virulence genes associated with APEC are often located on IncF plasmids, named ColV plasmids because their ability to code for production of colicin V, a small protein from the microcine family
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Fertility genes enable a plasmid's genetic information to be transferred from a donor to a recipient strain.
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Cells transformed with a plasmid vector only were used as negative controls.
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We constructed plasmid fusions by cloning the promoter regions of the wild-type parent and the inversion mutant onto the pRS550 plasmid.
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Plasmid-borne resistance to nourseothricin was observed not only in E. coli from pigs fed subtherapeutic doses of nourseothricin, but also in similar bacteria from employees at the pig farms.
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However, much antibiotic resistance seems to be acquired by the transfer of plasmids from other species of bacteria via conjugation, which of course does not explain the ultimate origin of the information.
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Asuragen is the preeminent partner for contract development and GMP manufacturing of RNA - and DNA-based standards and controls including plasmid DNA, Armored RNA®, and capped and uncapped IVT RNA.
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The labelled tyr T DNA fragments of interest were separated from the remainder of the plasmids on 6% polyacrylamide gels.
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These techniques range from simple calcium phosphate precipitation to sequestering the plasmid DNA in lipid vesicles that fuse to the cell membrane and deliver the contents to the cytoplasm (as illustrated in Figure 8).
Archive 2005-10-01
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Escherichia coli were transformed with the respective plasmids by chemical transformation.
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A plasmid containing this 3.1 kb of wild-type genomic DNA was cotransfected with a trp1 plasmid into protoplasts isolated from an auxotrophic thn1 mutant strain.
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Like bacteria, archaeans have no internal membranes and their DNA exists as a single loop called a plasmid.
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The GAACGACTCTACAATGTTGTtGTAGAtACAGAAGTTACTG GTAAAAAG oligonucleotide was used to mutagenize full-length SMC2 cDNA (a gift from K. Yokomori), which was subsequently inserted into NheI and XhoI sites of pcDNA3. 1, to produce the pcDNA3. 1/SMC2-2 complementation plasmid, resistant to RNAi with RNA duplex #2, due to silent mismatch mutation at the siRNA-corresponding site.
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We screened for loss-of-function mutations that would rescue a strain that contains three nonessential mutations affecting tubulin expression and that is inviable in the absence of a low-copy plasmid expressing tubulin.
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Instead I've ended up working 10-11ish hour days trying to play catch up to get the World's Most Uncooperative Plasmid to ligate my oligonucleotides and getting the yeast ready to do the shuffle.
Archive 2008-05-01
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Cells transfected with GFP and a plasmid encoding short hairpin RNA (shRNA) targeting cplx 2 were isolated by fluorescence-activated cell sorting and analyzed by immunoblotting.
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That vehicle (often a subcellular structure called a plasmid) is then inserted into a different cell, which activates the gene, leading to the production of a scientifically or commercially useful protein.
Scientists create cell based on man-made genetic instructions
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Fertility genes enable a plasmid's genetic information to be transferred from a donor to a recipient strain.
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Pps were produced by transfecting 293T cells with four plasmids: an HA expression plasmid, an NA expression plasmid, a Gag-pol-encoding plasmid, and a reporter plasmid.
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Alcohol (isopropanol) precipitation concentrates the eluted plasmid DNA, which is collected by centrifugation, washed, and redissolved in buffer.
Archive 2005-10-01
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Thus we were able to clone the DNA sequence that flanked the transposon insertion using plasmid rescue.
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Williams PH, Warner PJ (1980) ColV plasmid-mediated, colicin V-independent iron uptake system of invasive strains of
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By separating these viral coding sequences onto three separate plasmids, the probability of producing undesired replication-competent virus is very low.
The Scientist
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First, it has been observed that conjugative plasmids mediate gene transfer in various environments such as soil and rhizosphere, plant surfaces, water, or human gut.
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Many studies have shown that production of colicin V is not necessary for pathogenicity of E. coli by itself but it is a marker for other properties associated with the presence of ColV plasmid
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Objective To survey the antimicrobial resistance and plasmid profiles of Neisseria gonorrhoeae isolated from a hospital.
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However, as described above, this plasmid was found to exhibit wild type R-M.
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Once the gene of interest is ligated to the RFP plasmid, it can be cloned by inserting the pRFP into bacterial cells.
Archive 2005-10-01
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This phenomenon is similar to suicidal defense against phage infection, or phage exclusion, programmed by prophages and plasmids, and may well play a similar role.
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The inserts were ligated into the plasmid vector and primary transformants were generated as per the supplied protocol.
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For sporulation, diploid cells with plasmids were first grown in selective media.
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The plasmid DNA was then circularized with ligase and used to transform E. coli.
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Subcloning and comparative restriction enzyme analysis were carried out with the replication origin from tne integrated F' plasmid and that from the autonomous F plasmid.
