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Titre du document / Document title
Construction of a restriction-endonuclease-Eam I05I-generated T-vector for high-throughput cloning and expression
Auteur(s) / Author(s)
BAOLI WANG (1) ; HUI LIANG (2) ; RUI LIU (1) ; XIAOXIA LI (1 3) ; BEI SUN (1) ; RUI ZHANG (1) ; SHANYI GUO (1) ; GANG GUO (1) ; JINGYU ZHANG (1) ; CHENLIN DAI (4) ;
Affiliation(s) du ou des auteurs / Author(s) Affiliation(s)
(1) Key Research Laboratory of Hormone and Development Affiliated to the Ministry of Health, Institute of Endocrinology, Tianjin Medical University, Tianjin 300070, CHINE
(2) Department of Medical Diagnosis, Tianjin Medical University, Tianjin 300070, CHINE
(3) College of Basic Medical Science, Tianjin Medical University, Tianjin 300070, CHINE
(4) Tianjin Medical University Hospital, Tianjin 300070, CHINE
Résumé / Abstract
A novel T-vector was constructed that could be used for direct cloning and expression of PCR-amplified cDNA. The technique was based on the insertion into the parent vector of two endonuclease-Eam I 1051 restriction sequences spaced by an expression cassette of the full-length β-galactosidase, which helped to improve cloning efficiency and to minimize the non-recombinant background of the T-vector when used to clone PCR products. Moreover, this method took advantage of the reconstitution of the rarest restriction sequence of Mssl to enable directional cloning. These advantages make the T-vector suitable for high-throughput expression and analysis.
Revue / Journal Title
Biotechnology and applied biochemistry ISSN 0885-4513 CODEN BABIEC
Source / Source
2007, vol. 48 (1), pp. 29-33 [5 page(s) (article)] (12 ref.)
Langue / Language
Anglais
Editeur / Publisher
Portland Press, London, ROYAUME-UNI (1986) (Revue)
Mots-clés anglais / English Keywords
Hydrolases ; Glycosidases ; O-Glycosidases ; Enzyme ; Polymerase chain reaction ; β-Galactosidase ; Vector ; Restriction endonuclease ;
Mots-clés français / French Keywords
Hydrolases ; Glycosidases ; O-Glycosidases ; Enzyme ; Réaction chaîne polymérase ; β-Galactosidase ; Vecteur ; Restriction endonuclease ;
Mots-clés espagnols / Spanish Keywords
Hydrolases ; Glycosidases ; O-Glycosidases ; Enzima ; Reacción cadena polimerasa ; β-Galactosidase ; Vector ; Restriction endonuclease ;
Mots-clés d'auteur / Author Keywords
directional cloning ; β-galactosidase; high-throughput expression ; case chain reaction (PCR) ; T-vector. polymerase chain reaction (PCR) ; T-vector ;
Localisation / Location
INIST-CNRS, Cote INIST : 17979, 35400016082084.0040
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