Targeting vectors for gene diversification by meiotic recombination in Neurospora crassa.
We have constructed a pair of vectors, pDV2 and pDV3, that enable targeted insertion of exogenous DNA into Linkage Group I of Neurospora crassa at the his-3 locus. Transplaced sequences are inserted between his-3 and the cog(L) recombination hot spot and include his-3 mutations that allow meiotic recombination initiated by cog(L) to be monitored. Selection of correctly placed transforming DNA is based on complementation between different his-3 alleles borne by the plasmids and transformation hosts. The system allows investigation of the effect of any given sequence on recombination as well as diversification of sets of related sequences in vivo for directed evolution of genes.
Rasmussen
J Paul
JP
School of Biological Sciences, Adelaide, South Australia, 5001, Australia.
Bowring
Frederick J
FJ
Yeadon
P Jane
PJ
Catcheside
David E A
DE
D001483:Base Sequence; D003001:Cloning, Molecular; D004269:DNA, Bacterial; D018390:Gene Targeting; D005816:Genetic Complementation Test; D005818:Genetic Engineering; D005822:Genetic Vectors; D006639:Histidine; D007074:Immunoglobulin G / Q000235:genetics; D007145:Immunoglobulin kappa-Chains; D008540:Meiosis; D008969:Molecular Sequence Data; D009492:Neurospora crassa / Q000235:genetics* / Q000502:physiology; D011995:Recombination, Genetic
D004269:DNA, Bacterial; D007074:Immunoglobulin G; D007145:Immunoglobulin kappa-Chains; D006639:Histidine