pIZ/V5-His Vector Kit昆虫表达载体系统V800001 BioVector NTCC质粒载体菌种细胞基因保藏中心

pIZ/V5-His Vector Kit昆虫表达载体系统V800001

The InsectSelect™ System allows constitutive stable or transient expression of your
protein of interest in insect cell lines. The system utilizes a single expression
vector, pIZ/V5-His to express your gene of interest. This 2.9 kb vector has the
following features:
• OpIE2 promoter for high-level, constitutive expression of the gene of interest
(Theilmann and Stewart, 1992)
• Zeocin™ resistance gene for selection of stable cell lines (Hegedus et al., 1998;
Pfeifer et al., 1997)
• Optional C-terminal peptide containing the V5 epitope and 6xHis tag for
detection and purification of your protein of interest

Description of
System
The gene of interest is cloned into pIZ/V5-His and transfected into Sf9 or High
Five™ cells using lipid-mediated transfection. After transfection, cells can be
assayed for expression of the gene of interest. Once you have confirmed that
your gene expresses, you can select for a stable polyclonal population or stable
clonal cell lines using Zeocin™ as a selection agent. Stable cell lines can be used
to express the protein of interest in either adherent culture or suspension
culture.

Description of
Promoter
Baculovirus immediate-early promoters utilize the host cell transcription
machinery and do not require viral factors for activation. The OpIE2 promoter
is from the baculovirus Orgyia pseudotsugata multicapsid nuclear polyhedrosis
virus (OpMNPV). The virus’ natural host is the Douglas fir tussock moth;
however, the promoter allows protein expression in Lymantria dispar (LD652Y),
Spodoptera frugiperda cells (Sf9) (Hegedus et al., 1998; Pfeifer et al., 1997), Sf21
(Invitrogen), Trichoplusia ni (High Five™) (Invitrogen), Drosophila (Kc1, SL2)
(Hegedus et al., 1998; Pfeifer et al., 1997), and mosquito cell lines (unpublished
data). The OpIE2 promoter has been sequenced and analyzed. For more
detailed information on this promoter, see page 26.

Expression Levels The OpIE2 promoter provides relatively high levels of constitutive expression,
although not all proteins will be expressed at levels equivalent to those obtained
from baculovirus very late promoters (e.g. polyhedrin or p10). However, other
proteins may be expressed more efficiently in the InsectSelect™ System than in
baculovirus systems (Jarvis et al., 1996). To date, expression levels range from 1
μg/ml (human IL-6; Invitrogen) to 8–10 μg/ml (human melanotransferrin)


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