Site-directed mutagenesis

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Site-Directed Mutagenesis Study

Structural elements of the rat 1.t-opioid receptor important in ligand receptor binding and selectivity were examined using a site-directed mutagenesis approach. Five single amino acid mutations were made, three that altered conserved residues in the ~ 8, and K receptors (Asn 15°t Ala, His297 to Ala, and Tyr326 to Phe) and wo de igned to test for ~i/8 selectivity (lie198 to Val and Va1202 to li...

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Construction of a recombinant vector for site-directed mutagenesis in Salmonella typhimurium

BACKGROUND: Among all common techniques in sitedirectedmutagenesis, λ Red recombinase system has beenwidely used to knock out chromosomal genes in bacteria. In thismethod, there is always the risk of DNA Linear digestion byhost's restriction enzymes that leads to the low frequency ofrecombination. OBJECTIVES:To overcome this, we constructeda recombinant vector to disrupt phoP gene in Salmonella...

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Serratia marcescens B4A chitinase thermostability enhancement by S390I QuikChange site directed mutagenesis

Thermostable chitinases are useful for industrial and biotechnological applications. This paper reports the stabilization of chitinase from Serratia marcescens B4A through rational mutagenesis. Changing of Ser 390 to Ile in S. marcescens. The stabilization was enhanced through entropic stabilization by reduction of the loop length and also by increasing of the beta chain length. With this repla...

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Homemade site directed mutagenesis of whole plasmids.

Site directed mutagenesis of whole plasmids is a simple way to create slightly different variations of an original plasmid. With this method the cloned target gene can be altered by substitution, deletion or insertion of a few bases directly into a plasmid. It works by simply amplifying the whole plasmid, in a non PCR-based thermocycling reaction. During the reaction mutagenic primers, carrying...

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Site-directed mutagenesis of large plasmids.

A protocol combining recombination PCR and long-distance PCR is demonstrated to be highly accurate and rapid for site-directed mutagenesis of large (> 10 kb) plasmids. Application of this protocol to the generation of mutant rabies virus glycoproteins expressed by the baculovirus/insect cell system illustrates the usefulness of this approach in facilitating structure-function relationships in t...

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ژورنال

عنوان ژورنال: Biochemical Journal

سال: 1986

ISSN: 0264-6021,1470-8728

DOI: 10.1042/bj2370001