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  • genes
  • By removing the genes in the plasmid that caused the tumor and adding in novel genes, researchers were able to infect plants with A. tumefaciens and let the bacteria insert their chosen DNA into the genomes of the plants. (wikipedia.org)
  • The genes necessary to degrade oil were carried on plasmids, which could be transferred among species. (wikipedia.org)
  • By irradiating the transformed organism with UV light after plasmid transfer, Prof. Chakrabarty discovered a method for genetic cross-linking that fixed all four plasmid genes in place and produced a new, stable, bacterial species (now called Pseudomonas putida) capable of consuming oil one or two orders of magnitude faster than the previous four strains of oil-eating microbes. (wikipedia.org)
  • The hok/sok system involves three genes: hok, host killing - a long lived (half-life 20 minutes) toxin sok, suppression of killing - a short lived (half-life 30 seconds) RNA antitoxin mok, modulation of killing - required for hok translation When E. coli undergoes cell division, the two daughter cells inherit the long-lived hok toxin from the parent cell. (wikipedia.org)
  • organism
  • The use of E. coli as the experimental organism has allowed many generations and large populations to be studied in a relatively short period of time. (wikipedia.org)
  • Moreover, due to the long use of E .coli as a principle model organism in molecular biology, a wide array of tools, protocols, and procedures were available for studying changes at the genetic, phenotypic, and physiological levels. (wikipedia.org)
  • Ananda Mohan Chakrabarty (Bengali: আনন্দমোহন চক্রবর্তী Ānandamōhan Cakrabartī), Ph.D. is a Bengali American microbiologist, scientist, and researcher, most notable for his work in directed evolution and his role in developing a genetically engineered organism using plasmid transfer while working at GE, the patent for which led to landmark Supreme Court case, Diamond v. Chakrabarty. (wikipedia.org)
  • Cloning
  • Often these methods involve one or more cloning steps, which can be time-consuming, particularly when the study involves the construction of many plasmid derivatives. (asm.org)
  • For reason of ease and convenience, cloning is often performed using E. coli. (wikipedia.org)
  • The discovery of artificially induced competence in E. coli created an efficient and convenient procedure for transforming bacteria which allows for simpler molecular cloning methods in biotechnology and research, and it is now a routinely used laboratory procedure. (wikipedia.org)
  • bacteria
  • Lenski chose to carry out the experiment with the bacteria grown in a glucose-limited minimal medium called DM25, which was initially developed by Bernard Davis for use in isolating auxotrophic mutants of E. coli using penicillin as a selective agent. (wikipedia.org)
  • The first type I toxin-antitoxin system to be found in gram-positive bacteria is the RNAI-RNAII system of the pAD1 plasmid in Enterococcus faecalis. (wikipedia.org)
  • phage
  • At the upper left, the plasmid and the phage are shown as concentric circles with the phage outside. (asm.org)
  • However, in 1970, Morton Mandel and Akiko Higa showed that E. coli may be induced to take up DNA from bacteriophage λ without the use of helper phage after treatment with calcium chloride solution. (wikipedia.org)
  • recombinant
  • Manipulation of recombinant DNA, which is almost exclusively performed using the host E. coli, constitutes one of the fundamental methodologies of molecular biotechnology. (springer.com)
  • By demonstrating the utility of shuttle phasmids as DNA transporters between E. coli plasmids and mycobacteriophages, this paved the way for recombinant DNA research for mycobacteria. (wikipedia.org)
  • proteins
  • Fusion proteins are produced in E.coli usually in an insoluble form which facilitates purification.Hybrid proteins larger than 90 kDa are not expressed as efficiently as shorter ones. (stanford.edu)
  • However, not all proteins can be successfully expressed in E. coli, or be expressed with the correct form of post-translational modifications such as glycosylations, and other systems may therefore be used. (wikipedia.org)
  • The general goal of the experiment was to study the response of dehydrated and metabolically dormant microorganisms (spores of Bacillus subtilis, cells of Deinococcus radiodurans, conidial spores of Aspergillus species) and cellular constituents (plasmid DNA, proteins, purple membranes, amino acids, urea) to the extremely dehydrating conditions of outer space, in some cases in combination with irradiation by solar UV light. (wikipedia.org)
  • stably
  • Some DNA, however, cannot be stably maintained in E. coli, for example very large DNA fragments, and other organisms such as yeast may be used. (wikipedia.org)
  • protein
  • Permits positive selection for inserts, based on the dominance of the wild-type rpsL (ribosomal protein S12) gene in the plasmid. (stanford.edu)
  • The high level of gene expression from such plasmids is often desirable, for example in the production of large amounts of a protein for purification purposes. (asm.org)
  • chromosome
  • To avoid such complications, a variety of methods for integrating DNA from plasmids into the chromosome have been employed. (asm.org)
  • RNAI
  • This rate of degradation is aided by the pcnB (plasmid copy number B) gene product, which polyadenylates the 3' end of RNAI targeting it for degradation by PNPase. (wikipedia.org)
  • fragment
  • The BglII-HindIII fragment (nt 1392 of trpE to the end of the trpD sequence present in pKRS101) was replaced with a BamHI-EcoRI fragment and an EcoRI-HindIII fragment, both from the MCS of M13mp12 to make plasmid pATH1 (see GenBank M32985). (stanford.edu)
  • The SmaI-SmaI fragment from the MCS of pATH1 was deleted and the remaining plasmid religated to make plasmid pATH2 (see GenBank M33624). (stanford.edu)
  • growth
  • Growth of cells - E. coli cells are more susceptible to be made competent when it is growing rapidly, cells are therefore normally harvested in the early log phase of cell growth when preparing competent cells. (wikipedia.org)
  • The most striking adaptation reported so far is the evolution of aerobic growth on citrate, which is unusual in E. coli, in one population at some point between generations 31,000 and 31,500. (wikipedia.org)
  • strain
  • In E. coli strain K-12 there are four long direct repeats (ldr) which encode short open reading frames of 35 codons organised in a homologous manner to the hok/sok system. (wikipedia.org)
  • copy
  • Replicates to high copy number in E.coli and Enterobacter aerogenes. (stanford.edu)
  • However, there are also disadvantages to the use of high-copy-number plasmids. (asm.org)
  • In addition, heterogeneity of plasmid copy number within a population can result in a variation in expression level among single cells. (asm.org)
  • Daughter cells without a copy of the R1 plasmid die because they do not have the means to produce more sok antitoxin transcript to inhibit translation of the inherited hok mRNA. (wikipedia.org)
  • cell
  • The killing system is said to be postsegregational (PSK), since cell death occurs after segregation of the plasmid. (wikipedia.org)
  • larger
  • Assembled by F. Pfeiffer, MPI Biochemie, Martinsried E.coli transformed with this plasmid produce larger colonies when an insert is present in the polylinker than when there is no insert. (stanford.edu)
  • sequence
  • There are 11 DnaA binding sites/boxes on the E. coli origin of replication out of which three boxes R1, R2 and R4 (which have a highly conserved 9 bp consensus sequence 5' - TTATC/ACACA ) are high affinity DnaA boxes. (wikipedia.org)
  • Malgorzata Adamczyk and Grazyna Jagura-Burdzy: "Spread and survival of promiscuous IncP-1 plasmids", Acta Biochimica Polonica, Vol 50, no. 2/2003, p. 425-453 LEWIS C. INGRAM, M. H. RICHMOND, AND R. B. SYKES: "Molecular Characterization of the R Factors Implicated in the Carbenicillin Resistance of a Sequence of Pseudomonas aeruginosa Strains Isolated from Burns", ANTIMICROBIAL AGENTS AND CHEMOTHERAPY, Feb. 1973, p. 279-288 Thomas CM et al. (wikipedia.org)
  • During conjugation, the rolling circle mode of replication starts at the oriT ('T' for transfer) sequence of the FAT plasmid. (wikipedia.org)
  • strains
  • The role and the epidemiology of non-traditional enteropathogenic E. coli serogroup strains are not well established. (uzh.ch)
  • E. coli O157:H45 EPEC strains, however, are described in association with enterocolitis and sporadic diarrhea in human. (uzh.ch)
  • During a previous study on the prevalence of E. coli O157 in healthy cattle in Switzerland, E. coli O157:H45 strains originating from 6 fattening cattle and 5 cows were isolated. (uzh.ch)
  • This is especially interesting for strains such as P. putida, which can be host to a variety of catabolic conjugal plasmids (Sevastsyanovich et al. (micrornainhibitors.com)
  • EPEC
  • Moreover, a large outbreak associated with E. coli O157:H45 EPEC was reported in Japan in 1998. (uzh.ch)
  • protein
  • If a "phagemid" vector is used (a simplified display construct vector) phage particles will not be released from the E. coli cells until they are infected with helper phage, which enables packaging of the phage DNA and assembly of the mature virions with the relevant protein fragment as part of their outer coat on either the minor (pIII) or major (pVIII) coat protein. (wikipedia.org)
  • Recently an actin-like protein has been found in a gram-positive bacterium Bacillus thuringiensis, which forms a microtubule-like structure and is involved in plasmid segregation. (wikipedia.org)
  • Because the plasmid is the "vehicle" from which the immunogen is expressed, optimising vector design for maximal protein expression is essential. (wikipedia.org)
  • Alkaline phosphatase in E. coli is located in the periplasmic space and can thus be released using techniques that weaken the cell wall and release the protein. (wikipedia.org)
  • proteins
  • Alkaline phosphatase in E. coli is uncommonly soluble and active within elevated temperature conditions such as 80 °C. Due to the kinetic energy induced by this temperature the weak hydrogen bonds and hydrophobic interactions of common proteins become degraded and therefore coalesce and precipitate. (wikipedia.org)
  • replicate
  • The RK2 Plasmid is a broad-host-range plasmid belonging to the incP incompatibility group It is notable for its ability to replicate in a wide variety of single-celled organisms, which makes it suitable as a genetic engineering tool. (wikipedia.org)
  • structure
  • J A Kornacki, C H Chang, and D H Figurski: "kil-kor regulon of promiscuous plasmid RK2: structure, products, and regulation of two operons that constitute the kilE locus. (wikipedia.org)
  • The precise structure and function of the isozyme in E.coli is solely geared to supply a source of inorganic phosphate when the environment lacks this metabolite. (wikipedia.org)
  • another
  • Try to use a positive control (ask some college to give you another ampR plasmid) , you mention something but it is not clear if is working or not. (protocol-online.org)
  • RK2 may sometimes be referred to as pRK2, which is also the name of another, unrelated plasmid. (wikipedia.org)
  • contains
  • While the outer membrane of E. coli contains porins that are permeable to phosphorylated compounds, the inner membrane does not. (wikipedia.org)
  • Various
  • Various virulence factors (stx, eae, ehxA, astA, EAF plasmid, bfp) of different categories of pathogenic E. coli were screened by different PCR systems. (uzh.ch)