| Transpose-Ad™ Adenoviral Vector System
The Transpose-Ad™ Adenoviral Vector System is an E. coli
based system, taking advantage of the site-specific transposition
of the transposon Tn7. The gene of interest is cloned into
an MCS lying on a mini Tn7, flanked by both IRs (Inverted
Repeats) at both extremities of transposon Tn7. The recombined
transfer vector is subsequently introduced into E. coli
cells (HighQ-1 Transpose-Ad™ 294 cells) harboring a plasmid,
Transpose-Ad™ 294, that carries the Adenovirus backbone.
The same cells also carry a helper plasmid expressing the
transposase. The transposase ensures the insertion of the gene
of interest into its target site, attTn7, strategically located in the
lacZ gene on the Transpose-Ad™ 294 plasmid. Positive
recombinants are scored as β-gal- colonies on plates containing
Bluo-gal and IPTG.
White colonies are picked and the purified recombined plasmid
is introduced in chemically competent E. coli cells
(HighQ-1™) for amplification. The recombinant Transpose-
Ad™ 294 plasmid is extracted from the cells and then linearized
with Pac I before being transfected into human QBIHEK
293A cells, where the recombinant Adenovirus will be
generated and amplified.
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Advantages
- No plaque purification necessary
- Uses site-specific transposition, a highly efficient means of recombination
- Rapid and convenient blue/white color screening method, no
restriction enzyme mapping necessary
- No electroporation required
- The adenoviral backbone used for transposition is already contained
in the cells where transposition takes place and does not
require an additional transformation
Ordering Information
The Basic Kit contains the following components:
pCR259 transfer vector
pCR276 transfer vector
HighQ-1 Transpose-Ad™ 294 cells
HighQ-1™ cells
QBI-Infect viral particles
Transfection reagents (2M CaCl2, 0.1X TE and 2X HBS)
Products sold separately
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