New lambda and plasmid vectors for expression cloning in mammalian cells.
This report describes the construction of a new family of lambda phage and plasmid cloning vectors. lambda GDST3/T7 allows cDNA insertion up to 14 kb; it is derived from lambda NM1151 by the insertion of a multiple cloning site containing eight unique restriction sites. The two asymmetrical SfiI sites are flanked by the T3 and T7 promoters for direct sequencing and in vitro transcription/translation. The same multiple cloning site is also present in both orientations in the eukaryotic expression plasmids, pGDSV3 and pGDSV7. By exploiting the superior discrimination of the signal-to-noise ratio of the lambda vectors for primary screening (by either nucleic acids or antibody probes), relevant cDNAs can thus be efficiently transferred through SfiI sites into the plasmids pGDSV3/7 for functional secondary screening by expression in mammalian cells.
Del Sal
G
G
Laboratorio Nazionale Consorzio Interuniversitario per le Biotecnologie, Trieste, Italy.
Manfioletti
G
G
Gustincich
S
S
Ruaro
E
E
Schneider
C
C
D000818:Animals; D010582:Bacteriophage lambda / Q000235:genetics*; D001483:Base Sequence; D003001:Cloning, Molecular; D005822:Genetic Vectors; D008969:Molecular Sequence Data; D010957:Plasmids