A common multiple cloning site in a set of vectors for expression of eukaryotic genes in mammalian, insect and bacterial cells.
Here, we describe the construction of plasmid vectors facilitating expression of cloned genes in bacteria and in cells of mammalian and insect origin. Two types of multiple cloning site (MCS) were designed based on the MCS in the expression vector lambda gt11Sfi-Not. In the first set of vectors a start Met codon was included in the same reading frame as in lambda gt11Sfi-Not to support expression of partial cDNA clones. Thus a cDNA insert of lambda gt11Sfi-Not could be shuttled among the new vectors for expression. The other set of vectors without a start codon were suitable for expression of cDNA carrying their own start Met codon. By Western blot analysis and by transactivation of a reporter plasmid in co-transfections we show that cDNA is very efficiently expressed in NIH 3T3 cells under control of the elongation factor 1 alpha promoter.
Pallisgaard
N
N
Department of Molecular Biology, Aarhus University, Denmark.
Pedersen
F S
FS
Birkelund
S
S
Jørgensen
P
P
D016475:3T3 Cells; D017398:Alternative Splicing; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D051792:Basic Helix-Loop-Helix Proteins; D002460:Cell Line; D015500:Chloramphenicol O-Acetyltransferase / Q000096:biosynthesis / Q000378:metabolism; D003001:Cloning, Molecular / Q000379:methods*; D004268:DNA-Binding Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D004926:Escherichia coli; D005822:Genetic Vectors; D018257:Helix-Loop-Helix Motifs; D007313:Insecta; D008322:Mammals; D051379:Mice; D008969:Molecular Sequence Data; D009036:Moths; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification; D015183:Restriction Mapping; D014157:Transcription Factors / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D014162:Transfection
D051792:Basic Helix-Loop-Helix Proteins; D015500:Chloramphenicol O-Acetyltransferase; D004268:DNA-Binding Proteins; D011994:Recombinant Proteins; D014157:Transcription Factors; C489532:Tcf12 protein, mouse