Invitrogen Bac to Bac Expression System
Invitrogen Bac to Bac Expression System
Invitrogen Bac to Bac Expression System
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Map and Features of pFast<strong>Bac</strong> HT, continued<br />
Features of the<br />
Vec<strong>to</strong>r<br />
The pFast<strong>Bac</strong> HT A (4856 bp), B (4857 bp), and C (4858 bp) vec<strong>to</strong>rs contain the<br />
following elements. All features have been functionally tested.<br />
Feature<br />
Polyhedrin promoter (P PH )<br />
6xHis tag<br />
TEV recognition site<br />
Multiple cloning site<br />
SV40 polyadenylation signal<br />
Tn7L and Tn7R<br />
f1 origin<br />
Ampicillin resistance gene<br />
pUC origin<br />
Gentamicin resistance gene<br />
Benefit<br />
Allows efficient, high-level expression of your<br />
recombinant protein in insect cells (O'Reilly et<br />
al., 1992).<br />
Allows purification of your recombinant<br />
protein using a metal-chelating resin such as<br />
ProBond (Catalog no. K850-01) or Ni-NTA<br />
(Catalog no. K950-01).<br />
Permits removal of the N-terminal tag from<br />
your recombinant protein using TEV protease<br />
(Carring<strong>to</strong>n and Dougherty, 1988; Dougherty et<br />
al., 1988).<br />
Allows restriction enzyme-mediated cloning of<br />
your gene of interest.<br />
Permits efficient transcription termination and<br />
polyadenylation of mRNA (Westwood et al.,<br />
1993).<br />
Mini Tn7 elements that permit site-specific<br />
transposition of the gene of interest in<strong>to</strong> the<br />
baculovirus genome (i.e. bmon14272 bacmid)<br />
(Luckow et al., 1993).<br />
Allows rescue of single-stranded DNA.<br />
Allows selection of the plasmid in E. coli.<br />
Permits high-copy replication and maintenance<br />
in E. coli.<br />
Permits selection of the recombinant bacmid in<br />
DH10<strong>Bac</strong> E. coli.<br />
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