| Product |
pGEM-Z
Vectors |
| Description |
The pGEM-Z vectors are designed for use
as general purpose cloning vectors as well as for the
highly efficient synthesis of RNA in vitro. The vectors
contain both the SP6 and T7 RNA polymerase promoters flanking
the multiple cloning regions. The vectors also contain
the lac Z æ-peptide and a unique multiple cloning
site arrangement which allows for the color selection
for recombinants. This arrangement gives rise to a functional
æ-peptide which is capable of complementing the
product of the lac Z delta M 15 gene to produce functional
ß-galactosidase. Cells containing the lac Z delta
M 15 gene on an F' and also containing a pGEM-Z vector
will be blue in color when plated on indicator media containing
IPTG and X-Gal. However, when the lac æ-peptide
is disrupted by cloning into the pGEM-Z multiple cloning
region, complementation does not occur and ß-galactosidase
activity is produced. Therefore, bacterial colonies harboring
recombinant pGEM-Z vector constructs remain white.
The pGEM-3Z and pGEM-4Z vectors were the first vectors
to incorporate a color screening for recombinants and
also contain the SP6/T7 RNA polymerase promoters. These
vectors are differentiated by the orientation of their
multiple cloning region. Other vectors include the origin
of replication of the filamentous bacteriophage f1. For
induction of single-stranded DNA (ssDNA), bacterial cells
containing pGEM-Zf recombinants are infected with an appropriate
helper phage. The plasmid then enters the f1 replication
mode and the resulting ssDNA is exported from the cell
as an encapsidated virus-like particle. The single-stranded
plasmid DNA is purified from the supernatant by simple
precipitation and extraction procedures. The orientation
of the f1 (either + or -) determines which of the strands
of the plasmid will be secreted.
Additional vectors include the forementioned features
and unique multiple cloning regions designed for specific
applications. The pGEM-5Z (+/ -) and pGEM-7Z (+/ -) vectors
were constructed for use with the Erase-a-base system
for the generation of deletions using exonuclease III.
The multiple cloning region of these vectors contain cleavage
sites for restriction enzymes that produce 5' overhangs
or blunt ends (sensitive to exonuclease III). These sites
are flanked on both sides by blocks of restriction sites
that generate 3' overhangs (resistant to exonuclease III).
The pGEM-9Zf (-) vector include certain restriction sites
(i.e., Sfi EcoRI, and NotI ) that can be utilized in direct
cDNA cloning strategies or for convenient sub-cloning
of cDNAs cloned into the Lambda gt 11 Sfi-Not or Lambda
gt22 vectors.
All of pGEM-Z vectors are available separately. |
| Storage |
-20°C |
| Storage
Buffer |
10mM Tris-HCl, pH 7.5
and 1mM EDTA |
| Host |
E.coli HB101 |
|
