منابع مشابه
Site-Directed Mutagenesis Using Oligonucleotide-Based Recombineering
Methods enabling mutational analysis of distinct chromosomal locations, like site-directed mutagenesis, insertion of foreign sequences or in-frame deletions, have become of fast growing interest since complete bacterial genome sequences became available. Various approaches have been described to modify any nucleotide(s) in almost any manner. Some genetic engineering technologies do not rely on ...
متن کاملSite-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...
متن کاملCloning independent site-directed mutagenesis using total RNA as template.
Several site-directed mutagenesis methods have been developed and are being currently used in research. All these methods apply three or four primers, including one or two bearing the appropriate mutation, and most use PCR to amplify the desired sequence and to incorporate the mutation (1–6). Although several developments have improved the robustness and reliability of the method, all of them r...
متن کاملSite-directed mutagenesis using altered beta-lactamase specificity.
Promega's new GeneEditorTM in vitro Site-Directed Mutagenesis System* provides a high efficiency, reliable procedure for the generation and selection of oligonucleotide-directed mutants. The Selection Oligonucleotides provided in this system encode mutations which alter the substrate specificity of beta-lactamase, the gene encoding ampicillin resistance, commonly found on cloning vectors. This ...
متن کاملConstruction 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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ژورنال
عنوان ژورنال: BIO-PROTOCOL
سال: 2011
ISSN: 2331-8325
DOI: 10.21769/bioprotoc.29