Table 2.

Primers used for the site-directed mutagenesis

The underlined letters indicate the changes in nucleotide sequence. The bold letters indicate restriction sites.

Name	Sequence 5′ → 3′
Insertion into pRS416-pTPI
pRS-78TMEM-Fa	GTATCTAGAATGCATACTAACAAAGA (XbaI)
pRS-TMEM-Ra2	CATCTCGAGTTAAAAACCTG (XhoI)
pRS-E108G-F	TATTGTATCTGGACTTGGAG
pRS-E108G-R	CTCCAAGTCCAGATACAATA
pRS-R126H-F	GATATAATCATTTGACAGTTCTTG
pRS-R126H-R	CAAGAACTGTCAAATGATTATATC
pRS-R126C-F	GATATAATTGTTTGACAGTTCTTG
pRS-R126C-R	CAAGAACTGTCAAACAATTATATC
pRS-G304R-F	GTAACCATTATTAGAGGAATTG
pRS-G304R-R	CAATTCCTCTAATAATGGTTAC
Insertion into pNZ8048
Ins-F-4	AGCTCAGCCATGGGTCATC (NcoI)
Ins-R-1	CTCTTACCTTGAAGCTTTCA (HindIII)
E108G-F	TATTGTGTCCGGATTAGGTG
E108G-R	CACCTAATCCGGACACAATA
R126H-F	CTATAACCACCTGACCGTG
R126H-R	CACGGTCAGGTGGTTATAGC
R126C-F	CTATAACTGCCTGACCGTG
R126C-R	CACGGTCAGGCAGTTATAGC
G304R-F	ACCATCATTAGAGGAATCG
G304R-R	CGATTCCTCTAATGATGGTGAC