site-directed mutagenesis
A structure-based site-directed mutagenesis study on the neurolysin (EC 3.4.24.16) and thimet oligopeptidase (EC 3.4.24.15) catalysis
... The QuikChange site-directed mutagenesis kit (Stratagene, La Jolla, CA, USA) was used to introduce speci¢c point mutations in the wild type EP24.15 or EP24.16 cDNA, as previously described [11]. ...
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Identification of residues involved in nucleotidyltransferase activity of JHP933 from helicobacter pyloriby site-directed mutagenesis
... active site has been ...active site and little is known about the functional mechanism of ...rational site-directed mutagenesis combined with enzymatic activity assays, we confirm the ...
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ISSN 0100-879X BIOMEDICAL SCIENCES AND CLINICAL INVESTIGATION
... by site-directed mutagenesis, expressed in bacteria and purified using affinity ...containing site-directed mutations at positions Q39 and E451 was used to demonstrate that the effect ...
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Heterologous Proteins Production in Escherichia coli: An Investigation on the Effect of Codon Usage and Expression Host Optimization
... by site-directed mutagenesis and the expression level of the recombinant protein is analyzed by means of sodium dodecyl sulfate polyacry- lamide gel electrophoresis (SDS-PAGE) and Western ...
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Development of constructs for recombinant expression of human follicle-stimulating hormone receptor in rod cells of the zebrafish
... XIII Figure 17: PCR product inserted in the the pcDNA3.1/V5-His-TOPO © vector with the topoisomerase covalently bound to it (pcDNA3.1/V5-His © TOPO ® TA Expression Kit, 2004). ...
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Initiation of RNA Polymerization and Polymerase Encapsidation by a Small dsRNA Virus.
... active site (Fig 1A). Through site-directed mutagenesis, we determined that the loop structure most likely functions as a priming platform to support the binding of a single priming nucleotide ...
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Construction of the mutants by PCR mutagenesis
... Research was further stimulated after the crystal structures of α-AI1 complexed with PPA (Bompard- Gilles et al., 1996), Tenebrio molitor α-amylase (TMA) alone (Strobl et al., 1998), and in complex with α-AI1 (Nahoum et ...
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Characterization and regulation of a bacterial sugar phosphatase of the haloalkanoate dehalogenase superfamily, AraL, from Bacillus subtilis
... start site) was amplified from chromosomal DNA of the ...start site of the operon) was amplified from the wild-type strain with oligo- nucleotides ARA253 and ARA477 (Table 1), which carry unique XbaI and ...
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In-frame amber stop codon replacement mutagenesis for the directed evolution of proteins containing non-canonical amino acids: identification of residues open to bio-orthogonal modification.
... rational site-directed mutagenesis: limited understanding of the effect of a nAA on protein structure and function leading to the use of a fairly rudimentary design ...particular site may not ...
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Dissection of the TssB-TssC interface during type VI secretion sheath complex formation.
... Leu, Ile) while the opposite face is composed of polar or positively charged residues (Arg, Gln). The charge partition in the C- terminal region of the helix is better balanced but we observed a leucine-rich face, on the ...
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Membrane docking geometry of GRP1 PH domain bound to a target lipid bilayer: an EPR site-directed spin-labeling and relaxation study.
... The previously described optimized, fully functional Cysless GRP1 human PH domain (residues 255–392 and C293S/C327A/ C343S) was employed as the background for creation of a single- Cys mutant library [26,27]. Single ...
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Novel enzyme-polymer conjugates for biotechnological applications
... B. thermocatenulatus lipase (Genkbank number X95309) mutants were prepared as pre- viously described (Romero et al., 2012). Briefly, all site-directed mutagenesis experiments were carried out by PCR ...
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Intergenic incompatibilities reduce fitness in hybrids of extremely closely related bacteriophages
... known site of contact between the coat protein and the external scaffolding protein (D) during the procapsid stage of assembly (Dokland et ...by site-directed mutagenesis by substituting this ...
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Site-directed mutation of a laccase from Thermus thermophilus: Effect on the activity profile
... he QuikChange site-directed mutagenesis kit was purchased from Stratagene (Santa Clara, USA). Mo- lecular weight markers were from New England Bi- oLabs (Ipswich, USA). Primers were synthesized by ...
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Functional Domains of the Bacillus subtilis Transcription Factor AraR and Identification of Amino Acids Important for Nucleoprotein Complex Assembly and Effector Binding†
... random mutagenesis of the araR gene and site-directed mutagenesis guided by molec- ular modeling techniques to identify the residues within AraR that are involved in DNA binding, dimer ...
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Structural insights into cellulolytic and chitinolytic enzymes revealing crucial residues of insect β-N-acetyl-D-hexosaminidase.
... overall topology as well as amino acid sequence shares very little similarity (Fig. 1) [8]. Insect OfHex1 from the Asian corn borer (Ostrinia furnacalis) is a b-N-acetyl-D-hexosaminidase specialized for chitin ...
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Purification, cloning, characterization and essential amino acid residues analysis of a new ι-carrageenase from Cellulophaga sp. QY3.
... subsites. Site-directed mutagenesis followed by kinetic analysis identified three strictly conserved residues at subsites 21 and +1 of i-carrageenases, G228, Y229 and R254 in CgiA_Ce, which played ...
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Defining the functional domain of programmed cell death 10 through its interactions with phosphatidylinositol-3,4,5-trisphosphate.
... and site-directed mutagenesis, it appears that PDCD10 is a six helical bundle protein composed of a two heptad repeat, and a5 is an important amphipathic helix housing five lysine residues essential ...
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A sucrose binding protein homologue from soybean exhibits GTP-binding activity that functions independently of sucrose transport activity
... by site-directed mutagenesis the GTP binding site of S64/SBP2 to this imperfect A consensus ...binding site (Thr283Leu and Lys284Glu changes) do not represent structurally neutral ...
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Medicine and the Task of Healing
... practical site where the knowledge of science is applied in the service of ethics, and where ethics encounters the conditions of ...is directed towards overcoming pathology—but pathology understood in the ...
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