Open AccessDevelopment of host and vector for high‐efficiency transformation and gene disruption in Debaryomyces hansenii
Author(s) -
Minhas Anupriya,
Biswas Dipanwita,
Mondal Alok K.
Publication year - 2009
Publication title -
fems yeast research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.991
H-Index - 92
eISSN - 1567-1364
pISSN - 1567-1356
DOI - 10.1111/j.1567-1364.2008.00457.x
Subject(s) - debaryomyces hansenii , biology , transformation (genetics) , selectable marker , gene , genetics , halotolerance , auxotrophy , shuttle vector , homologous recombination , vector (molecular biology) , computational biology , yeast , bacteria , recombinant dna , mutant
Debaryomyces hansenii is one of the most osmotolerant and halotolerant yeasts. The molecular mechanisms underlying its extreme osmotolerance and halotolerance have drawn considerable attention in the recent past. However, progress in this regard has been limited due to lack of availability of a transformation system and molecular tools to study the functions of the genes in D. hansenii . Here, we have described the development of an efficient transformation system for D. hansenii that is based on a histidine auxotrophic recipient strain and the DhHIS4 gene as the selectable marker. By screening the D. hansenii genomic library, we have isolated several autonomous replication sequences that can be used for constructing a replicating vector. Moreover, our study is the first to demonstrate gene disruption in D. hansenii by homologous recombination.