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From a Short Amino Acidic Sequence to the Complete Gene

AuthorsMiñambres Rodríguez, Baltasar ; Olivera, Elías R.; García, Belén; Naharro, Germán; Luengo, José M.
Issue Date2000
CitationBiochemical and Biophysical Research Communications 272(2): 477-479 (2000)
AbstractA useful strategy directed to the isolation of a required gene with a high GC content is reported. Using a degenerate oligonucleotide probe, deduced from the amino terminus of a protein, it is possible to obtain a fragment of DNA containing its encoding gene by PCR amplification. Furthermore, the cloning of a desired gene can be accomplished in two steps by using an oligonucleotide deduced (i) from an internal sequence, (ii) from a consensus sequence, or (iii) from a DNA sequence adjacent to a disrupting element (transposon, insertion sequence, cassette). This method, which could be applied to a bacteriophage, plasmid, or cosmid genomic library, has been successfully used for cloning several genes from different biological systems.
Description3 pages, 1 figure.-- PMID: 10833438 [PubMed].
Publisher version (URL)http://dx.doi.org/10.1006/bbrc.2000.2709
Appears in Collections:(IGM) Artículos
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