Site-directed mutagenesis in Brucella abortus S19 by overlap extension PCR-based procedure
نویسندگان
چکیده
منابع مشابه
Combined Overlap Extension PCR Method for Improved Site Directed Mutagenesis
The combined overlap extension PCR (COE-PCR) method developed in this work combines the strengths of the overlap extension PCR (OE-PCR) method with the speed and ease of the asymmetrical overlap extension (AOE-PCR) method. This combined method allows up to 6 base pairs to be mutated at a time and requires a total of 40-45 PCR cycles. A total of eight mutagenesis experiments were successfully ca...
متن کاملconstruction of mutant wbka gene in brucella abortus s19 by overlap extension pcr
background: causing site direct mutation can be one of the efficient methods to evaluate the characteristics and properties of various genes. brucellosis is the most common zoonotic infectious disease that would cause great economic losses. thus, recognition of pathogenic and immunogenic factors in the genus brucella can lead to control this health problem. objectives: considering the importanc...
متن کاملA rapid and efficient method for site-directed mutagenesis using one-step overlap extension PCR.
A rapid method is described to efficiently perform site-directed mutagenesis based on overlap extension polymerase chain reaction (OE-PCR). Two template DNA molecules in different orientations relative to only one universal primer were amplified in parallel. By choosing a high dilution of mutagenic primers it was possible to run an overlap extension PCR in only one reaction without purification...
متن کاملSite-directed mutagenesis by overlap extension using the polymerase chain reaction.
Overlap extension represents a new approach to genetic engineering. Complementary oligodeoxyribonucleotide (oligo) primers and the polymerase chain reaction are used to generate two DNA fragments having overlapping ends. These fragments are combined in a subsequent 'fusion' reaction in which the overlapping ends anneal, allowing the 3' overlap of each strand to serve as a primer for the 3' exte...
متن کاملModification of a PCR-based site-directed mutagenesis method.
Site-directed mutagenesis is a powerful tool for producing mutants to assess the importance of specific amino acid residues in a protein’s structure and/or function. We wanted to generate mutants of human ETS1 cDNA in the pET15b vector (Novagen, Madison, WI, USA) from which we had been producing wild-type protein for structural studies (11). Since we were subject to the constraints of this vect...
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ژورنال
عنوان ژورنال: Journal of the Hellenic Veterinary Medical Society
سال: 2018
ISSN: 2585-3724,1792-2720
DOI: 10.12681/jhvms.15468