CaSpeR5, a family of Drosophila transgenesis and shuttle vectors with improved multiple cloning sites

摘要

The pCaSpeR (1) and pUAST vectors (2) are two of the most commonly used Drosophila transformation vectors. However, although they have great utility in their current form, their multiple cloning sequences (MCSs) have a limited number of unique restriction sites (Figure 1). This is particularly true for the pUAST vector, whose MCS has only five unique sites. Further, neither of the MCSs in pCaSpeR or pUAST are present in small shuttle or cloning vectors, which is problematic, because the large size (>8 kb) of the transgenesis vectors requires sequence manipulations such as site-directed mutagenesis or deletion dropouts to be done in small plasmid vectors and the modified DNA to be moved to the transgenesis vectors. The lack of matching shuttle vectors further constrains the usable cloning sites and can make moving large genomic fragments between a cloning vector and a transgenesis vector problematic.