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Summary: 5451TECHNICAL PAPER RESEARCH REPORT
INTRODUCTION
The ability to engineer genetically modified organisms is essential
for establishing the function of genes during development, disease,
homeostasis, repair and regeneration (Gama Sosa et al., 2010;
Ristevski, 2005). However, a crucial step in engineering genetically
modified organisms is the design and generation of the transgene
DNA constructions required for a given experiment. For the past
thirty years, DNA constructions have been created primarily
through restriction enzyme digestion and ligation. However,
cloning with restriction enzymes becomes progressively more
cumbersome as the complexity of the engineered constructs
increases. For this reason, a site-specific recombination-based
DNA cloning method was developed that circumvents the use of
restriction enzymes (Hartley et al., 2000). The advent of
recombination-based cloning brought a series of diverse and
pioneering studies showing the utility of this technology in creating
DNA constructions for transgenesis (Fisher et al., 2006; Hope et
al., 2004; Ikeya et al., 2005; Kappas et al., 2008; Kwan et al., 2007;
Nyabi et al., 2009; Semple et al., 2010; Skarnes et al., 2011).
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