A Modified Unique Site Elimination Mutagenesis in Constructing a Chloramphenicol Resistance-Encoding pGEM® Vector
نویسندگان
چکیده
منابع مشابه
A modified unique site elimination mutagenesis in constructing a chloramphenicol resistance-encoding pGEM vector.
Recombinant DNA research requires new and versatile cloning vectors. Many of the popular plasmid vectors, such as the pUC (4), pTZ (Amersham, Arlington Heights, IL, USA) and pGEM (Promega, Madison, WI, USA) series, share common characteristics, including high copy number, sizes of around 3 kbp and the presence of a multiple cloning sequence (MCS) within the coding sequence for a part of the la...
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...
متن کاملSite-directed mutagenesis of multi-copy-number plasmids: Red/ET recombination and unique restriction site elimination.
Existing methods for site-directed plasmid mutagenesis are restrained by the small spectrum of modifications that can be introduced by mutagenic primers and the amplicon size limitations of in vitro DNA synthesis. As demonstrated here, the combined use of Red/ET recombination and unique restriction site elimination enables extensive manipulation regardless of plasmid size and DNA sequence eleme...
متن کاملMethod of site-directed mutagenesis using long primer-unique site elimination and exonuclease III.
troducing undesired mutations during the preparation of the vector, can negatively affect the frequency of gene targeting in ES cells (13). In summary, the method described here offers an attractive alternative for the preparation of replacement vectors for gene targeting. This protocol can be used to perform site-directed mutagenesis in long genomic sequences cloned into plasmids with the high...
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...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
ژورنال
عنوان ژورنال: BioTechniques
سال: 1996
ISSN: 0736-6205,1940-9818
DOI: 10.2144/96206bm10