TABLE 1.

Oligonucleotides used for site-directed mutagenesis of TM XI

Mutated nucleotides are underlined, restriction sites are bold. Restriction sites deleted are indicated in parentheses. Oligonucleotides used for cysteine scanning mutagenesis are shown. All mutations were to Cys.

Mutation	Oligonucleotide sequence	Restriction site
Gln449	5′-GTATCGTGAAGCTTACCCCCAAGGACTGCTTCATCATCGCC-3′	HindIII
Phe450	5′-CCCCCAAGGACCAATGCATCATCGCCTATGG-3′	SstI
Ile451	5′-CCCAAGGACCAGTTCTGCATCGCATATGGGGGCCTGCGAG-3′	NdeI
Ile452	5′-GGACCAGTTCATCTGCGCATATGGGGGCCTGC-3′	FspI
Ala453	5′-CCAGTTCATCATCTGCTATGGAGGCCTGCGAGGGGC-3′	StuI
Tyr454	5′-CAGTTCATCATCGCATGCGGGGGCCTGCGAGG-3′	SphI
Gly455	5′-CATCATCGCCTATTGCGGCTTAAGAGGGGCCATCGCC-3′	AflII
Gly456	5′-CATCGCCTATGGGTGCTTAAGAGGGGCCATCGCC-3′	AflII
Leu457	5′-GCCTATGGGGGCTGCCGCGGGGCCATCGCCTTC-3′	SacII
Arg458	5′-CATCGCCTATGGAGGCCTGTGTGGGGCCATCGCC-3′	StuI
Gly459	5′-GGGGGCCTGCGATGCGCAATCGCCTTCTCTC-3′	FspI
Ala460	5′-GGGGCCTGCGAGGATGCATCGCCTTCTCTC-3′	NsiI
Ile461	5′-GGCCTGCGAGGGGCATGCGCCTTCTCTCTGGG-3′	SphI
Ala462	5′-CTGCGAGGGGCCATATGCTTCTCTCTGGGC-3′	NdeI
Phe463	5′-GAGGGGCCATCGCATGCTCTCTGGGCTACC-3′	SphI
Ser464	5′-GGCCATCGCCTTCTGCCTAGGCTACCTCCTGG-3′	AvrII
Leu465	5′-CATCGCCTTCTCTTGCGGGTACCTCCTGGACAAG-3′	KpnI
Gly466	5′-CGCCTTCTCTCTGTGCTACCTTCTAGA CAAGAAGCACTTC-3′	XbaI
Tyr467	5′-CCTTCTCTCTGGGCTGTCTACTGGACAAGAAGCAC-3′	AccI
Leu468	5′-CTCTCTGGGCTACTGTCTAGACAAGAAGCACTTC-3′	XbaI
Leu469	5′-GCCTTCTCTCTGGGATATCTCTGC GACAAGAAGCACTTC-3′	EcoRV
Asp470	5′-GGGCTACCTCCTGTGCAAAAAGCACTTCCCCATG-3′	(Xmn I)