Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
1
pubmed:dateCreated
1989-8-10
pubmed:abstractText
A simple and efficient site-specific mutagenesis method is described. First, a single-stranded (ss) circular vector is linearized at the site where the desired mutation will be introduced. To do this, an oligodeoxynucleotide complementary to the target region of the ss vector and containing a restriction enzyme recognition sequence is annealed to the circular ss vector, and the partial double-strand formed is subsequently cleaved with that enzyme. Then, another oligodeoxynucleotide spanning the nick and carrying the mutation is annealed to the linearized ss DNA template and the annealed mixture is used directly to transform Escherichia coli without prior enzymatic DNA synthesis in vitro. The procedure has been applied successfully to constructing insertion, deletion, and point mutations in both M13 phage vectors and plasmid vectors containing the f1 origin of replication.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Feb
pubmed:issn
0003-2697
pubmed:author
pubmed:issnType
Print
pubmed:day
15
pubmed:volume
177
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
120-4
pubmed:dateRevised
2006-4-21
pubmed:meshHeading
pubmed:year
1989
pubmed:articleTitle
Oligodeoxynucleotide-directed cleavage and repair of a single-stranded vector: a method of site-specific mutagenesis.
pubmed:affiliation
Institut Jacques Monod, CNRS, Paris, France.
pubmed:publicationType
Journal Article