pvitro1-nmcs plasmid vector哺乳動物雙基因組成型高效表達(dá)載體質(zhì)粒

GENERAL PRODUCT USE
pVITRO is a family of plasmids developed mainly for in vitro studies.
They allow the ubiquitous and constitutive co-expression of two genes
of interest. pVITRO plasmids can be stably transfected in mammalian
cells and the genes of interest are expressed at high levels. Each
pVITRO plasmid is available with either two multiple cloning sites or
two reporter genes.
pVITRO1-nmcs plasmid is selectable with kanamycin in E. coli and
G418 in mammalian cells. It contains two multiple cloning sites (MCS)
for the convenient cloning of two cDNAs.
PLASMID FEATURES
? rEF1 and mEF1 prom: pVITRO1-nmcs plasmid carries two
elongation factor 1 alpha (EF-1α) promoters, from rat and mouse
origins. Similarly to their human counterpart1
, both promoters display a
strong activity that yield similar levels of expression. EF-1α promoters
are expressed at high levels in all cell cycles and lower levels during G0
phase. EF-1α promoters are also non-tissue specific; they are highly
expressed in all cell types.
? SV40 enhancer which is comprised of a 72-base-pair repeat allows
the enhancement of gene expression in a large host range2
. The
enhancement varies from 2-fold in non-permissive cells to 20-fold
in permissive cells.
? CMV enhancer: The major immediate early enhancer of the human
cytomegalovirus (HCMV), located between nucleotides -118 and -524,
is composed of unique and repeated sequence motifs. The HCMV
enhancer can substitute for the 72-bp repeats of SV40 and is severalfold
more active than the SV40 enhancer3
.
? SV40 pAn: the Simian Virus 40 late polyadenylation signal enables
efficient cleavage and polyadenylation reactions resulting in high
levels of steady-state mRNA. The efficiency of this signal was first
described by Carswell et al.4
? pMB1 ori: a minimal E. coli origin of replication to limit vector size,
but with the same activity as the longer Ori.
? FMDV IRES: The internal ribosome entry site of the Foot and Mouth
Disease Virus enables the translation of two open reading frames from
one mRNA with high levels of expression5
.
? EM7 is a bacterial promoter that enables the constitutive expression
of the antibiotic resistance gene in E. coli.
? Neo: The neo gene from Tn5 confers resistance to Kanamycin in E.
coli and G418 in mammalian cells. In bacteria, neo is expressed from
the constitutive E. coli EM7 promoter. In mammalian cells, neo is
transcribed from the rat EF-1α promoter as a polycistronic mRNA and
translated via the FMDV IRES.
? EF1 pAn is a strong polyadenylation signal. InvivoGen uses a
sequence starting after the stop codon of the EF1 cDNA and finishing
after a bent structure rich in GT.
? MCS1 and MCS2: To facilitate cloning each multiple cloning site contains
several restriction sites that are compatible with many other enzymes

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