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Titolo:
A MODIFIED OVERLAP EXTENSION PCR METHOD TO CREATE CHIMERIC GENES IN THE ABSENCE OF RESTRICTION ENZYMES
Autore:
WURCH T; LESTIENNE F; PAUWELS PJ;
Indirizzi:
CTR RECH PIERRE FABRE,DEPT CELLULAR & MOL BIOL,17 AVE JEAN MOULIN F-81106 CASTRES FRANCE
Titolo Testata:
Biotechnology techniques
fascicolo: 9, volume: 12, anno: 1998,
pagine: 653 - 657
SICI:
0951-208X(1998)12:9<653:AMOEPM>2.0.ZU;2-5
Fonte:
ISI
Lingua:
ENG
Soggetto:
POLYMERASE-CHAIN-REACTION; MUTAGENESIS;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Science Citation Index Expanded
Science Citation Index Expanded
Citazioni:
11
Recensione:
Indirizzi per estratti:
Citazione:
T. Wurch et al., "A MODIFIED OVERLAP EXTENSION PCR METHOD TO CREATE CHIMERIC GENES IN THE ABSENCE OF RESTRICTION ENZYMES", Biotechnology techniques, 12(9), 1998, pp. 653-657

Abstract

A modified overlap extension technique for the creation of chimeric genes is described: the method consists in three PCR steps. The first step is a conventional PCR reaction, in which oligonucleotide primers are partially complementary at their 5' ends to the adjacent fragments that are fused to create the chimer. The second PCR step consists in the fusion of the PCR fragments generated in the first step using the complementary extremities of the primers. The third step corresponds tothe PCR amplification of the fusion product. The final PCR product isa chimeric gene built up with the different amplified PCR fragments. The technique is illustrated by the construction of a chimeric 5-hydroxytryptamine (5-HT, serotonin)(1B/D) receptor by combining one part ofthe human 5-HT1B (h5-HT1B) and two parts of the h5-HT1D receptor gene. The chimeric gene expressed in Cos-7 cells yielded similar binding properties as the wild type h5-HT1D receptor.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 05/04/20 alle ore 18:54:33