StarGate® Combinatorial Cloning
To complement Source BioScience LifeSciences's every increasing
portfolio of cDNA resources, we are pleased to announce the
distribution of a high quality combinatorial cloning system,
StarGate®. Any gene of interest can be systematically
transferred into E. coli, yeast, baculovirus and mammalian
expression systems. To enhance your experiment further, optimal
promoters, signal sequences and tag(s) may also be incorporated
into your final expression construct.
Introduction
The StarGate® cloning system uses efficient
procedures to generate functional recombinant proteins or protein
complexes which are of key importance in state-of-the-art life
sciences. Many tools including various expression hosts (bacteria,
yeast, insect and mammalian cells), promoters, affinity or
fluorescent tags are currently available to express, purify, detect
or immobilize recombinant proteins.
Due to the diverse nature of proteins, however, it is impossible
to predict which combination of these tools will perform best.
Therefore StarGate® offers rapid and highly efficient
sub-cloning of any gene - initially cloned into a Donor Vector - to
simultaneously fuse it with many different genetic surroundings via
transfer into Acceptor Vectors to generate Destination Vectors. The
latter enable the efficient expression of your protein with various
features (e.g. different tags and different promoters) in different
hosts.
Standard StarGate® Procedure
Step 1: Entry reaction
In the first step, a gene of interest (GOI) is equipped at both
ends with combinatorial sites (4 bases) by PCR and is inserted into
an Entry Vector by a simple one-tube reaction. The opened Entry
Vector donates the recognition sites and provides an operative
linkage with the combinatorial sites for the highly specific
StarGate® gene transfer process (step 2).
Step 2: Transfer reaction
After sequence confirmation, the resulting Donor Vector is the
source for the highly parallel sub-cloning of the GOI by a second
simple one tube reaction into a multitude of Acceptor Vectors, each
providing a different genetic surrounding like host specific
promoters and different purification tags. The resulting
Destination Vectors are then transformed into the corresponding
host cells for further experiments.
Acceptor Vectors are can be provided on request.
For StarGate® newcomers! We've compiled a 50%
discount for the "StarGate® Newcomer" Set which provides
all products required to perform the entire StarGate®
protocol. Please remember to choose a number and type of Acceptor
Vectors according to your needs and order them separately.
Source BioScience LifeSciences are also offering a cloning
service using the StarGate® cloning system. If you would
like further information regarding this or would like to discuss
your requirements, please contact
us to discuss your requirements.