Abstract
We have developed a method to capture inserts from P1 and P1 artificial chromosome (PAC) clones into a yeast–bacteria shuttle vector by using recombinogenic targeting. We have engineered a vector, pPAC-ResQ, a derivative of pClasper, which was previously used to capture inserts from yeast artificial chromosome clones. pPAC-ResQ contains DNA fragments flanking the inserts in P1 and PAC vectors as recombinogenic ends. When linearized pPAC-ResQ vector and P1 or PAC DNA are cotransformed into yeast, recombination between the two leads to the transfer of inserts into pPAC-ResQ. pPAC-ResQ clones thus obtained can be further modified in yeast for functional analysis and shuttled toEscherichia colito produce large quantities of cloned DNA. This approach provides a rapid method to modify P1/PAC clones for functional analysis.
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