Open AccessCharacterization of DhKHA1 , a gene coding for a putative Na + transporter from Debaryomyces hansenii
Author(s) -
CarcíaSalcedo Raúl,
Montiel Vera,
Calero Fernando,
Ramos José
Publication year - 2007
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.1567-1364.2007.00258.x
Subject(s) - debaryomyces hansenii , biology , saccharomyces cerevisiae , gene , mutant , heterologous expression , coding region , gene expression , microbiology and biotechnology , green fluorescent protein , transporter , biochemistry , genetics , yeast , recombinant dna
The KHA1 gene from Debaryomyces hansenii has been identified and characterized by heterologous expression in Saccharomyces cerevisiae . The gene is orthologous to ScKHA1 , previously reported in S. cerevisiae , and on the basis of the deduced amino acid sequence, Dh Kha1p can be classified as an Na + /H + transporter. Reverse transcriptase (RT)‐PCR experiments indicated that the expression level of DhKHA1 was not dependent on high pH or on the presence of a high salt level in the growth medium. Overexpression of DhKHA1 in a salt‐sensitive S. cerevisiae mutant ( ena1‐4 nha1 kha1 ) rendered cells specifically more tolerant to Na + . In addition, internal K + and Na + measurements and experiments performed with green fluorescence protein (GFP)‐tagged Dh Kha1p indicated the intracellular localization of this protein when expressed in S. cerevisiae .