113 research outputs found
SHORT COMMUNICATION - Cre-loxP recombination vectors for promoter studies
For promoter studies the cloning, subcloning and transfer to different
plasmid vectors usually requires use of restriction enzymes and
ligation reactions. One obstacle is the nucleotide polymorphisms of
eukaryotic genomic DNA, which has the consequence that a sequence often
differs from published sequences. Therefore sequencing, rigorous
restriction enzyme analysis or introduction of suitable sites has to be
performed prior to cloning and subcloning. In addition, conventional
methods using restriction enzymes, insert purifications and ligations
is expensive and labour demanding. We have developed a fast, efficient
and inexpensive Cre recombinase-loxP based method, which allows cloning
of promoter regions and subcloning of these into a variety of vectors
in a restriction enzyme independent manner. We here demonstrate that
expression of a number of reporter genes and a therapeutic gene from
both a viral and 2 mammalian promoters cloned by this recombinase
method have activities comparable to conventionally cloned plasmids
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