• In the present study, we describe the cloning of the KHV genome as a stable and infectious bacterial artificial chromosome (BAC) clone that can be used to produce KHV recombinant strains. (nih.gov)
  • CopyControl™ BAC Cloning Kits* are complete kits containing all molecular biology reagents needed for constructing high-quality, large-insert Bacterial Artificial Chromosome (BAC) libraries. (bjzblg.com)
  • If eutomation makes it possible to follow changes in a densely-mapped bacterial chromosome subjected to a variety of mutational situations, the resul- ting insipht: into chrenosene mechantes will be extremely va tuapics axtension Gv Ghis appreneh to ehiuccosa: ml dpbrner i her irmortene conecaucnecs Vor witereiandine a vide rane oe eilitdes pacvunetoaac gor cecil in which * good biological mechanisms of genctic recombination are available. (nih.gov)
  • 27 « Glaser, Donald A. Privileged Cormunication * , 4) Genetic characterization of the chromosome terminus and the regulation of cell division in E. coli. (nih.gov)
  • This hypothesis has recently received some experimental support: (a) studies of cell division following DNA, RNA, and protein inhibition at the time of chromosome termination in synchronous cultures (16-18) in@icate that the specific replication of the lest 0.5% of the chromosome. (nih.gov)
  • All commonly used cloning vectors in molecular biology have key features necessary for their function, such as a suitable cloning site and selectable marker. (wikipedia.org)
  • However, the most commonly used cloning vectors include plasmids and bacteriophages (phage λ) beside all the other available vectors. (exama2z.in)
  • Cloning vectors in yeast include yeast artificial chromosomes (YACs). (wikipedia.org)
  • The different types of vectors available for cloning are plasmids, bacteriophages, bacterial artificial chromosomes (BACs), yeast artificial chromosomes (YACs) and mammalian artificial chromosomes (MACs). (exama2z.in)
  • There are many types of cloning vectors, but the most commonly used ones are genetically engineered plasmids. (wikipedia.org)
  • Cloning is generally first performed using Escherichia coli, and cloning vectors in E. coli include plasmids, bacteriophages (such as phage λ), cosmids, and bacterial artificial chromosomes (BACs). (wikipedia.org)
  • Thus, the cloning vectors used often have elements necessary for their propagation and maintenance in E. coli, such as a functional origin of replication (ori). (wikipedia.org)
  • All cloning vectors have features that allow a gene to be conveniently inserted into the vector or removed from it. (wikipedia.org)
  • citation needed] Other cloning vectors may use topoisomerase instead of ligase and cloning may be done more rapidly without the need for restriction digest of the vector or insert. (wikipedia.org)
  • The gene, once cloned into the cloning vector (called entry clone in this method), may be conveniently introduced into a variety of expression vectors by recombination. (wikipedia.org)
  • Reporter genes are used in some cloning vectors to facilitate the screening of successful clones by using features of these genes that allow successful clone to be easily identified. (wikipedia.org)
  • CopyControl pCC1BAC cloning vectors are Cloning-Ready: linearized, dephosphorylated, and highly purified. (bjzblg.com)
  • Vectors - to carry, maintain and replicate cloned gene in host cell. (exama2z.in)
  • The cloning vectors are limited to the size of insert that they can carry. (exama2z.in)
  • All cloning vectors are carrier DNA molecules. (exama2z.in)
  • Phage T1-resistant TransforMax™ EPI300™-T1 R Electrocompetent E. coli cells, or standard TransforMax™ EPI300™ Electrocompetent E. coli cells, which are required for induction of CopyControl BAC clones to 10-20 copies per cell, are available separately. (bjzblg.com)
  • Phage display technology was further developed and improved by groups at the Laboratory of Molecular Biology with Greg Winter and John McCafferty, The Scripps Research Institute with Richard Lerner and Carlos Barbas and the German Cancer Research Center with Frank Breitling and Stefan Dübel for display of proteins such as antibodies for therapeutic protein engineering. (wikipedia.org)
  • The phage gene and insert DNA hybrid is then inserted (a process known as "transduction") into E. coli bacterial cells such as TG1, SS320, ER2738, or XL1-Blue E. coli. (wikipedia.org)
  • 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)
  • Those that remain can be eluted, used to produce more phage (by bacterial infection with helper phage) and to produce a phage mixture that is enriched with relevant (i.e. binding) phage. (wikipedia.org)
  • Phage eluted in the final step can be used to infect a suitable bacterial host, from which the phagemids can be collected and the relevant DNA sequence excised and sequenced to identify the relevant, interacting proteins or protein fragments. (wikipedia.org)
  • citation needed] The use of a helper phage can be eliminated by using 'bacterial packaging cell line' technology. (wikipedia.org)
  • Applications of phage display technology include determination of interaction partners of a protein (which would be used as the immobilised phage "bait" with a DNA library consisting of all coding sequences of a cell, tissue or organism) so that the function or the mechanism of the function of that protein may be determined. (wikipedia.org)
  • The practice of phage therapy, which uses bacterial viruses (phages) to treat bacterial infections, has been around for almost a century. (wjgnet.com)
  • Current research on the use of phages and their lytic proteins against multidrug-resistant bacterial infections, suggests phage therapy has the potential to be used as either an alternative or a supplement to antibiotic treatments. (wjgnet.com)
  • To do this work it was necessary to develop a way to pull plasmids apart, glue segments of them back together again, then propagate and clone new combinations of plasmid genes in living cells. (nih.gov)
  • These investigations proved a link between bacterial DNA carried in plasmids and the formation of resistance-plasmid DNA. (nih.gov)
  • A necessary next step in studying the molecular biology of plasmids was the reintroduction of plasmid DNA molecules into bacteria. (nih.gov)
  • This insertion led to a BAC plasmid that was stably maintained in bacteria and was able to regenerate virions when permissive cells were transfected with the plasmid. (nih.gov)
  • The cloning vector may be DNA taken from a virus, the cell of a higher organism, or it may be the plasmid of a bacterium. (wikipedia.org)
  • Another kind of selectable marker allows for the positive selection of plasmid with cloned gene. (wikipedia.org)
  • The action of interferons (IFNs) on virus-infected cells and surrounding tissues elicits an antiviral state that is characterized by the expression and antiviral activity of IFN-stimulated genes. (nature.com)
  • Beginning with his invention and development of EST sequencing to identify sequences of large numbers of expressed genes, Dr. Venter went on to determine the structural sequence of the hereditary materials of other mammalian, insect, bacterial, and viral genomes. (cornell.edu)
  • DNA fragment containing the desired genes to be cloned. (exama2z.in)
  • Gene amplification, the increase in the copy number of a portion of the genome, is a common manifestation of genome instability in tumour cells and an important mechanism of oncogene activation as well as drug resistance, since it leads to over-expression of relevant genes. (biomedcentral.com)
  • The term "gene cloning," "DNA cloning," "molecular cloning," and "recombinant DNA technology" all refer to same technique. (exama2z.in)
  • Host cell- in which recombinant DNA can replicate. (exama2z.in)
  • A fragment of DNA, containing the gene to be cloned, is inserted into a suitable vector, to produce a recombinant DNA molecule. (exama2z.in)
  • When the host cell divides, copies of the recombinant DNA molecule are passed to the progeny and further vector replication takes place. (exama2z.in)
  • the gene carried by the recombinant molecule is now said to be cloned. (exama2z.in)
  • It must possess some marker gene such that it can be used for later identification of recombinant cell (usually an antibiotic resistance gene that is absent in the host cell). (exama2z.in)
  • The recombinant vector is transformed into suitable host cell mostly, a bacterial cell. (exama2z.in)
  • Similarly, virions with a wild-type revertant TK sequence (the FL BAC revertant strain) were produced by the cotransfection of cells with the BAC and a DNA fragment encoding the wild-type TK sequence. (nih.gov)
  • A cloning vector is a small piece of DNA 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)
  • Multiple cloning sites are sometimes used to ensure that the fragments are inserted in all three possible reading frames so that the cDNA fragment is translated in the proper frame. (wikipedia.org)
  • Standard molecular typing methods such as multilocus sequence typing, restriction fragment length polymorphism, and fingerprinting using variable number of tandem repeats have shown an apparent lack of genetic diversity of M. ulcerans within individual geographic regions, which is indicative of a clonal population structure. (cdc.gov)
  • Another method of cloning without the use of DNA digest and ligase is by DNA recombination, for example as used in the Gateway cloning system. (wikipedia.org)
  • This displayed the different peptides on the outer surfaces of the collection of viral clones, where the screening step of the process isolated the peptides with the highest binding affinity. (wikipedia.org)
  • In mammalian cells gene amplification is a common manifestation of genome instability promoted by DNA double-strand breaks (DSBs). (biomedcentral.com)
  • Different pathways, mainly controlling either the cell cycle in response to DNA damage or the repair of the damage itself, maintain genome stability in mammalian cells. (biomedcentral.com)
  • Mammalian cells repair DSBs through two main mechanisms: (1) homologous recombination (HR), which requires large regions of homology, and (2) non-homologous end joining (NHEJ), which does not require extended homologies [ 11 ]. (biomedcentral.com)
  • SYBR ® is a registered trademark of Molecular Probes, Inc. (bjzblg.com)
  • The vector and the foreign DNA may be treated with a restriction enzyme that cuts the DNA, and 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 by molecular ligation. (wikipedia.org)
  • Isolation of DNA [gene of interest fragments to be cloned. (exama2z.in)
  • The restriction sites may be further used for sub-cloning into another vector if necessary. (wikipedia.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)
  • A selectable marker is carried by the vector to allow the selection of positively transformed cells. (wikipedia.org)
  • Shuttle vector which is designed to be maintained in two different organisms may also require two selectable markers, although some selectable markers such as resistance to zeocin and hygromycin B are effective in different cell types. (wikipedia.org)
  • The vector acts as a vehicle that transports the gene into a host cell usually a bacterium, although other types of living cell can be used. (exama2z.in)
  • Within the host cell the vector multiplies, producing numerous identical copies not only of itself but also of the gene that it carries. (exama2z.in)
  • The production of exact copies of a particular gene or DNA sequence using genetic engineering techniques is called gene cloning. (exama2z.in)
  • The study of genetic diversity within bacterial species has provided information on aspects such as virulence ( 1 , 2 ), antimicrobial drug resistance ( 3 ), epidemiology, and microbial evolution ( 4 - 7 ). (cdc.gov)
  • Anchor sequences are molecular markers that are unique loci in genetic linkage maps of multiple species and are used in comparative genomics for cross-species mapping along co-linear genomic regions. (nih.gov)
  • In 2 NIH-supported studies, a fast, powerful technique called whole-exome sequencing provided a molecular diagnosis for about 1 in 4 people suspected of having a genetic disorder. (nih.gov)
  • Targeted ablation of exon 2 of the Avian Leukosis Virus-A (ALV-A)receptor gene in a chicken fibroblast cell line by CRISPR abrogates ALV-A infection. (usda.gov)
  • Interferons (IFNs) - the body's first line of antiviral defence - are cytokines that are secreted by host cells in response to virus infection. (nature.com)
  • The high frequency of isochromosome formation in cells deficient for RAD54 suggests that homologous recombination proteins might play a role in preventing rearrangements at the centromeres. (biomedcentral.com)
  • The fact that gene amplification has never been detected in cells of normal origin [ 3 , 4 ] suggests that either control mechanisms that prevent the occurrence of gene amplification are active (such as the p53-mediated damage-sensing pathway), or cells carrying gene amplifications do not survive. (biomedcentral.com)
  • Cloning is one method used for isolation and amplification of gene of interest. (exama2z.in)
  • Then, immediately before DNA purification, clones can be induced to 10-20 copies per cell within 2 hours by adding CopyControl Induction Solution to the culture, for higher yields of BAC DNA. (bjzblg.com)
  • This may be a multiple cloning site (MCS) or polylinker, which contains many unique restriction sites. (wikipedia.org)
  • The short, 2-hour induction of CopyControl BAC clones to high-copy number does not decrease their stability, based on analysis of Hin d III restriction patterns of a large number of induced versus uninduced clones that ranged in size up to ~200 kb. (bjzblg.com)
  • Our results showed that both RAD54 and DNA-PKcs deficient cells are hypersensitive to γ-irradiation and generate methotrexate resistant colonies at a higher frequency compared to the proficient cell lines. (biomedcentral.com)
  • blocking protein synthesis during this replication will block the subsequent cell division. (nih.gov)
  • Most types of cell can produce IFN-α and IFN-β, which are the best-characterized type I IFNs, whereas IFN-γ is produced only by certain cells of the immune system, including natural killer (NK) cells, CD4 + T helper 1 (T H 1) cells and CD8 + cytotoxic T cells. (nature.com)
  • To maintain stability, CopyControl BAC clones are grown at very low copy number during the initial cloning and screening processes. (bjzblg.com)
  • Safely obtain higher DNA yields while maintaining the stability of single-copy-number clones. (bjzblg.com)
  • The epidemiology and mode of transmission of M. ulcerans disease are not fully understood, partly because no molecular typing method with sufficiently high resolution for microepidemiologic analyses is available. (cdc.gov)
  • This may involve the use of a gene lethal to the host cells, such as barnase, Ccda, and the parD/parE toxins. (wikipedia.org)
  • This typically works by disrupting or removing the lethal gene during the cloning process, and unsuccessful clones where the lethal gene still remains intact would kill the host cells, therefore only successful clones are selected. (wikipedia.org)
  • After a large number of cell divisions, a colony, or clone, of identical host cells is produced. (exama2z.in)
  • Selection of transformed host cells and identification of the clone containing the gene of interest. (exama2z.in)
  • In addition, the analysis of the cytogenetic organization of the amplicons revealed that isochromosome formation is a prevalent mechanism responsible for copy number increase in RAD54 defective cells. (biomedcentral.com)
  • In 1988, Stephen Parmley and George Smith described biopanning for affinity selection and demonstrated that recursive rounds of selection could enrich for clones present at 1 in a billion or less. (wikipedia.org)
  • Estimate the size of the CopyControl BAC clones in as little as 4 hours without culture, Not I digests, or pulse-field gel electrophoresis. (bjzblg.com)
  • NHEJ is an error prone process that dominates during the G1 to early S phase of the cell cycle whereas HR is mainly used in the late S and G2 phases. (biomedcentral.com)
  • The global antiviral state of the cell involves cross-talk between IFN signalling and pathways that regulate apoptosis, inflammation and cellular stress-response programmes. (nature.com)
  • Cohen's DNA cloning research was the result of his interest in basic scientific inquiry into fundamental natural phenomena, not to create new tools for diagnosing human disease. (nih.gov)
  • We constructed human cell lines in which the expression of RAD54 and/or DNA-PKcs was constitutively knocked-down by RNA interference. (biomedcentral.com)
  • CopyControl BAC Cloning Kits reduce the time and labor required to construct a BAC library. (bjzblg.com)
  • Autoinduction Solution is added to the culture medium prior to inoculation, and also increases the cell growth leading to higher yield than standard Induction Solution. (bjzblg.com)
  • IFNs exert their biological functions by binding to specific cell-surface receptors. (nature.com)