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  • genome
  • Since 2013, the development of the CRISPR/Cas9 technology, based on a prokaryotic viral defense system, has also allowed for the editing of the genome, and mutagenesis may be performed in vivo with relative ease. (wikipedia.org)
  • These nucleases create site-specific double-strand breaks (DSBs) at desired locations in the genome. (wikipedia.org)
  • The primary advantage of this technique is its ability to eliminate any foreign DNA from the genome after the mutagenesis process. (wikipedia.org)
  • Since 2013, development of the CRISPR/Cas9 technology, based on a prokaryotic viral defense system, has also allowed for the editing or mutagenesis of the genome in vivo. (wikipedia.org)
  • enzyme
  • The coenzymes bind to the active site of an enzyme to promote catalysis. (wikipedia.org)
  • HindIII (pronounced "Hin D Three") is a type II site-specific deoxyribonuclease restriction enzyme isolated from Haemophilus influenzae that cleaves the DNA palindromic sequence AAGCTT in the presence of the cofactor Mg2+ via hydrolysis. (wikipedia.org)
  • The segment may be at an enzyme active site, or sequences that have structural significance or immunogenic property. (wikipedia.org)
  • reagents
  • Furthermore, if you want to alter the sequence at more than one site (e.g., changing several codons at one site, and adding a protein tag to the resulting mutant), you will need to perform multiple rounds of mutagenesis, and the cost of reagents and time will rise accordingly. (idtdna.com)
  • protein
  • Also called site-specific mutagenesis or oligonucleotide-directed mutagenesis, it is used for investigating the structure and biological activity of DNA, RNA, and protein molecules, and for protein engineering. (wikipedia.org)
  • The tertiary structure consists of a beta/alpha-barrel, with the coenzyme-binding site located at the carboxy-terminus end of the strands of the barrel.Alternative splicing of this gene results in two transcript variants encoding the same protein. (wikipedia.org)
  • The site-directed approach may be done systematically in such techniques as alanine scanning mutagenesis whereby residues are systematically mutated to alanine in order to identify residues important to the structure or function of a protein. (wikipedia.org)
  • specificity
  • Since the glycoprotein hormones contain a common α-subunit, and since both subunits are believed to participate in receptor binding (6, 7), interesting questions and models can be developed regarding subunit contact sites and hormone receptor contact sites, particularly those responsible for conferring specificity. (springer.com)
  • gene
  • There are numerous methods for achieving site-directed mutagenesis, but with decreasing costs of oligonucleotide synthesis, artificial gene synthesis is now occasionally used as an alternative to site-directed mutagenesis. (wikipedia.org)
  • The gene thus copied contains the mutated site, and is then introduced into a host cell as a vector and cloned. (wikipedia.org)
  • As was mentioned above, dsDNA gene fragments are compatible with familiar PCR mutagenesis methods, but they also offer some interesting advantages. (idtdna.com)
  • proteins
  • Highly conserved positions in the sequences of specific C1 oxidizing cellulose-active AA10 proteins were targeted for site-directed mutagenesis, and the C4 oxidizing activity of a MaLPMO10B variant, carrying two mutated residues, was almost completely lost. (bibsys.no)
  • experiments
  • As a result of the site-mutagenesis experiments previously outlined, it is thus proposed that Lys-125, Asp-123, and Asp-108 of HindIII function similarly to Lys-92, Asp-90, and Asp-74 in EcoRV, respectively. (wikipedia.org)
  • Analysis
  • Despite the lack of evidence suggesting an exact mechanism for the cleavage of DNA by HindIII, site-mutagenesis analysis coupled with more detailed studies of metal ion-mediated catalysis in EcoRV have led to the following proposed catalytic mechanism. (wikipedia.org)
  • homology
  • One of the known DSB repair pathways that are essentially functional in all organisms[citation needed] are the non-homologous end joining (NHEJ) and homology directed repair (HDR). (wikipedia.org)
  • molecules
  • The cofactor Mg2+ is believed to bind water molecules and carry them to the catalytic sites of the enzymes, among other cations. (wikipedia.org)
  • restriction
  • Despite the uncertainty concerning the structure-catalysis relationship of type II endonucleases, site-directed mutagenesis of the restriction endonuclease HindIII has provided much insight into the key amino acid residues involved. (wikipedia.org)
  • While restriction enzymes cleave at specific DNA sequences, they are first required to bind non-specifically with the DNA backbone before localizing to the restriction site. (wikipedia.org)
  • Type I restriction enzymes recognize specific sequences, but cleave DNA randomly at sites other than their recognition site whereas type II restriction enzymes cleave only at their specific recognition site. (wikipedia.org)
  • mouse
  • Insertional mutagenesis using transposons, retrovirus such as mouse mammary tumor virus and murine leukemia virus may be used to identify genes involved in carcinogenesis and to understand the biological pathways of specific cancer. (wikipedia.org)
  • enzymes
  • The structure of some enzymes are very well characterized, however, the function of some component of the active site is poorly understood. (wikipedia.org)
  • Gene
  • The previously identified core promoter region in the hTR gene has several features utilised by the basal RNA PolII transcription machinery, including one CCAAT-box and four Sp1 sites termed Sp1.1-Sp1.4. (biomedcentral.com)
  • Non-viral methods of gene delivery can be divided into transformation, where a cell incorporates foreign DNA from its surroundings, and conjugation, where gene transfer is a result of direct contact between two cells. (wikipedia.org)
  • The enzyme encoded by this gene is a member of the conserved DNA/RNA non-specific ββα-Me-finger nuclease family and possesses a unique site selectivity of poly(dG).poly(dC) sequences in double-stranded DNA. (wikipedia.org)
  • A plasmid expressing flippase (FLP) can be transformed into the recombined cells, which can specifically cleave FLP recognition target sites (FRTs) flanking the antibiotic resistance gene. (wikipedia.org)
  • In this case, a gene cassette with a dual selectable marker can be incorporated into the DNA at the specific location of mutagenesis. (wikipedia.org)