Phytochelatin synthase is required for tolerating metal toxicity in a basidiomycete yeast and is a conserved factor involved in metal homeostasis in fungi

Table 3 Primers used in this study

Name	Sequence (5′-3′)	Purpose
AS008	gtaacgccagggttttcccagtcacgacgCTGATTTCTGCGAAGAGG	Disruption of Sporobolomyces PCS1
AS002	GGTCTTTCCAGGGAGAGAGGGTTCATTCTGAGTGAG
AS003	CAAGTAGAACGAAGGGTTCGCATTCTCAACCATCTC
AS009	gcggataacaatttcacacaggaaacagcCAAGAGTTGTTCGGTGCG
AS005	GGGGTACCATGACGCTTGCCACCAAAGC	Cloning Sporobolomyces PCS1 cDNA
AS006	GCTCTAGACTAGTGACAACCCTTCTGC	Cloning Sporobolomyces PCS1 cDNA
ALID0562	TCTCTCCCTGGAAAGACC	Amplification of URA5 selectable marker
ALID0564	AACCCTTCGTTCTACTTG	Amplification of URA5 selectable marker
AS012	TCGGTTATCGGTTAAGGC	Gene replacement in S. pombe
AS013	GTCGACCTGCAGCGTACGTCCCTCTTGCAATGCTCG
AS014	CGAGCTCGAATTCATCGATTCCCAAAGGCGTTCTAG
AS015	TAAACGAAAGCACGAGCG
KanMX F	CGTACGCTGCAGGTCGAC
KanMX R	ATCGATGAATTCGAGCTCG
AS016	GCCCATATGACGCTTGCCACCAAAGC	Sporobolomyces PCS1 for expression in S. pombe
AS017	GCCCATATGCTAGTGACAACCCTTCTG	Sporobolomyces PCS1 for expression in S. pombe
AS018	CGGCATATGAACATTGTTAAACGAG	Amplifcation of S. pombe PCS1
AS019	ATTGGATCCTCACGTATTTTTACAGCAG	Amplifcation of S. pombe PCS1
ALID2091	CAAGCTGCTGTAAAAATACGATGGTGAGCAAGGGCGAG	GFP localization of S. pombe Pcs1
ALID2092	CTCGCCCTTGCTCACCATCGTATTTTTACAGCAGCTTG
AISV066	CCGGGATCCTTACTTGTACAGCTCGTC
AS004	CAAGAGTTGTTCGGTGCG	Wild type Sporobolomyces PCS1 for complementation
AS020	GGTCTTTCCAGGGAGAGACTGATTTCTGCGAAG	Wild type Sporobolomyces PCS1 for complementation
ALID1432	GAGTACATGGTCTACATG	GPD1 for northern blots
ALID1433	AAGGCAATGTGGCAGAGG	GPD1 for northern blots

Letters in italics are restriction enzyme cut sites introduced into the DNA sequences. Lower case letters indicate regions that were included for possible use for in vivo recombination into S. cerevisiae plasmids.

ISSN: 2054-3085