• 1986
  • There is a yeast shuttle vector (pWH5) that may cater for your needs that was published by Wright APH, Maundrell K, Heyer W-D, Beach D, Nurse P (1986) Vectors for the construction of gene banks and the integration of cloned genes in S.pombe and S.cerevisiae. (bio.net)
  • viral vector
  • The global viral vector and plasmid DNA manufacturing market is segmented on the basis of product, application, and geography. (medgadget.com)
  • Insertion of a vector into the target cell is usually called transformation for bacterial cells, transfection for eukaryotic cells, although insertion of a viral vector is often called transduction. (wikipedia.org)
  • For example, retroviruses leave a characteristic retroviral integration pattern after insertion that is detectable and indicates that the viral vector has incorporated into the host genome. (wikipedia.org)
  • ColE1
  • Early cloning experiments may be conducted using natural plasmids such the ColE1 and pSC101. (wikipedia.org)
  • For example, the ColE1 plasmid and its derivatives have the advantage of higher copy number and allow for chloramphenicol amplification of plasmid to produce a high yield of plasmid, however screening for immunity to colicin E1 is not technically simple. (wikipedia.org)
  • Another plasmid, RSF 2124, which is a derivative of ColE1, confers ampicillin resistance but is larger. (wikipedia.org)
  • sequence
  • 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)
  • Deposited by: Walter Messer Vector containing the attP sequence for integrating DNA into the lambda attachment site attB. (stanford.edu)
  • The target DNA sequence can be inserted into the vector in a specific direction if so desired. (wikipedia.org)
  • For example, prokaryotes expression vectors would have a Shine-Dalgarno sequence at its translation initiation site for the binding of ribosomes, while eukaryotes expression vectors would contain the Kozak consensus sequence. (wikipedia.org)
  • The vector itself is generally a DNA sequence that consists of an insert (transgene) and a larger sequence that serves as the "backbone" of the vector. (wikipedia.org)
  • Vectors designed specifically for the expression of the transgene in the target cell are called expression vectors, and generally have a promoter sequence that drives expression of the transgene. (wikipedia.org)
  • DNAs
  • In the last few years, several viral and nonviral (plasmid DNAs) have been standardized and optimized for this purpose. (medgadget.com)
  • circular
  • The "pJAZZ" linear vectors are shown to provide unprecedented ability to maintain regions of up to 30 kb that are unclonable in circular plasmids. (aacrjournals.org)
  • In this TOPO cloning method a linearized vector is activated by attaching topoisomerase I to its ends, and this "TOPO-activated" vector may then accept a PCR product by ligating both the 5' ends of the PCR product, releasing the topoisomerase and forming a circular vector in the process. (wikipedia.org)
  • Plasmids are small, closed-circular DNA molecules that exist from the chromosomes of their host. (wikipedia.org)
  • SmaI
  • 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)
  • An interim vector was constructed by inserting an EcoRI linker at the remaining SmaI site of pATH2. (stanford.edu)
  • ClaI
  • The order of the major features in this plasmid is: NotI - ampR - ClaI/MCS/EcoRI - attP - NotI - pMB1 ori - tetR. (stanford.edu)
  • personal communication) The order of the major features in this plasmid is:EcoRI - BglII - EcoRI - ClaI - HindIII - BamHI - rpsL - ori - bla. (stanford.edu)
  • molecule
  • In molecular cloning, a vector is a DNA molecule used as a vehicle to artificially carry foreign genetic material into another cell, where it can be replicated and/or expressed (e.g.- plasmid, cosmid, Lambda phages). (wikipedia.org)
  • A cloning vector is a DNA molecule that carries foreign DNA into a host cell, where it replicates, producing many copies of itself along with the foreign DNA. (wikipedia.org)
  • promoter
  • A larger number of mRNAs would express a greater amount of protein, and how many copies of mRNA are generated depends on the promoter used in the vector. (wikipedia.org)
  • ditags
  • The linkers on the outside of the ditags are cleaved with the enzyme that provided their binding and these sticky end ditags are concatenated randomly and placed into a cloning vector. (wikipedia.org)
  • Under intramolecular (dilute) conditions, vectors are re-circularized and ligated, leaving only the ditags in the vector. (wikipedia.org)
  • EcoRI
  • The EcoRI-HindIII fragment of MCS in this interim vector was replaced with the EcoRI-HindIII fragment containing the MCS of M13mp12. (stanford.edu)
  • fragment
  • Tha AvaII-AvaII fragment that spanned the PstI site in the bla gene of this interim vector was replaced with the corresponding AvaII fragment from pUC8, eliminating this PstI site, making the PstI site in the MCS unique, to form plasmid pATH11. (stanford.edu)
  • A cloning vector is a small piece of DNA, taken from a virus, a plasmid, or the cell of a higher organism, that can be stably maintained in an organism, and into which a foreign DNA fragment can be inserted for cloning purposes. (wikipedia.org)
  • After a DNA fragment has been cloned into a cloning vector, it may be further subcloned into another vector designed for more specific use. (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 vector is used to introduce a specific gene into a target cell, and can commandeer the cell's mechanism for protein synthesis to produce the protein encoded by the gene. (wikipedia.org)
  • The goal of a well-designed expression vector is the efficient production of protein, and this may be achieved by the production of significant amount of stable messenger RNA, which can then be translated into protein. (wikipedia.org)
  • There are differences in the machinery for protein synthesis between prokaryotes and eukaryotes, therefore the expression vectors must have the elements for expression that is appropriate for the chosen host. (wikipedia.org)
  • The promoters used in expression vector are normally inducible, meaning that protein synthesis is only initiated when required by the introduction of an inducer such as IPTG. (wikipedia.org)
  • Gene expression however may also be constitutive (i.e. protein is constantly expressed) in some expression vectors. (wikipedia.org)
  • Low level of constitutive protein synthesis may occur even in expression vectors with tightly controlled promoters. (wikipedia.org)
  • The expression vector is transformed or transfected into the host cell for protein synthesis. (wikipedia.org)
  • Different organisms may be used to express a gene's target protein, and the expression vector used will therefore have elements specific for use in the particular organism. (wikipedia.org)
  • All vectors may be used for cloning and are therefore cloning vectors, but there are also vectors designed specially for cloning, while others may be designed specifically for other purposes, such as transcription and protein expression. (wikipedia.org)
  • colonies
  • 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)
  • overhangs
  • DNA fragments thus generated contain either blunt ends or overhangs known as sticky ends, and vector DNA and foreign DNA with compatible ends can then be joined together by molecular ligation. (wikipedia.org)
  • genetic material
  • Such vectors have bacterial or viral elements which may be transferred to the non-bacterial host organism, however other vectors termed intragenic vectors have also been developed to avoid the transfer of any genetic material from an alien species. (wikipedia.org)
  • ligation
  • I suspect that the problem is with the gel purification and not with my digestions for the following reason: If I just do the digestion followed by the phosphatase (but without any gel purification), the kinase control works perfectly (the plasmid can recircularize), but unfortunately the negative control (ligation without kinase) produces approx. (bio.net)
  • Antibiotic
  • The plasmid pSC101, a natural plasmid from Salmonella panama, confers tetracycline resistance which allows for simpler screening process with antibiotic selection, but it is a low copy number plasmid which does not give a high yield of plasmid. (wikipedia.org)