Fast and easy method for construction of plasmid vectors using modified quick-change mutagenesis.
Plasmid vector construction is an essential step for molecular microbiology yet often a time-consuming process. Manipulation of the fungal genome to express genes to activate secondary metabolite production often requires creation of plasmid constructs in a reiterative fashion. Here we introduce a modified Quick-change site-directed mutagenesis method that allows for rapid and accurate construction of fungal transformation vectors.
Department of Medical Microbiology and Immunology, University of Wisconsin-Madison, Madison, WI, USA. jwbok@facstaff.wisc.edu
Jin Woo
JW
Bok
Nancy P
NP
Keller
D001233:Aspergillus nidulans / Q000235:genetics; D017931:DNA Primers / Q000235:genetics; D004271:DNA, Fungal / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D015252:Deoxyribonucleases, Type II Site-Specific / Q000378:metabolism; D005822:Genetic Vectors / Q000235:genetics*; D016681:Genome, Fungal / Q000235:genetics; D016297:Mutagenesis, Site-Directed / Q000379:methods*; D010957:Plasmids / Q000235:genetics*; D016133:Polymerase Chain Reaction; D013997:Time Factors; D014170:Transformation, Genetic
D017931:DNA Primers; D004271:DNA, Fungal; C020102:endodeoxyribonuclease DpnI; D015252:Deoxyribonucleases, Type II Site-Specific