Table 2.

Groups B and C: Primer sequences and template plasmids used for site-direct mutagenesis.

Mutation(s)	Primer sets	Primer location	Template Plasmid	Clone Number
N368Q	5′-CCTGCCCCGACAGATCACTGAGG-3′	1509-31	64-5	192-9
	5′-CCTCAGTGATCTGTCGGGGCAGG-3′		71-28	459-1
T370Q	5′-CCCCGAAATATCCAGGAGGGTGAAGCC-3′	1513-39	64-5	196-34,36
	5′-GGCTTCACCCTCCTGGATATTTCGGGG-3′		71-28	459-5
T352Q	5′-GAGGAGGACATCCAGATCCAGACGGTGACCTG-3′	1459-1490	71-28	457-24
	5′-CAGGTCACCGTCTGGATCTGGATGTCCTCCTC-3′
T352Q-T357Q	5′-CATCCAGACGGTGCAGTGGCCTGACGTG-3′	1473-1500	457-24	460-6
	5′-CACGTCAGGCCACTGCACCGTCTGGATG-3′
T352Q-T355Q-T357Q	5′-CATCCAGATCCAGCAGGTGCAGTGGCCTG-3′ 5′-CAGGCCACTGCACCTGCTGGATCTGGATG-3′	1467-95	460-6	461-4
T352V	5′-GAGGAGGACATCGTCATCCAGACGGTG-3′	1459-1490	71-28	458-3
	5′-CACCGTCTGGATGACGATGTCCTCCTC-3′
T352V-T357V	5′-CATCCAGACGGTGGTCTGGCCTGACGTG-3′	1473-1500	458-3	459-24
	5′-CACGTCAGGCCAGACCACCGTCTGGATG-3′
T352V-T355V-T357V	5′-CATCACCATCCAGGTGGTGACCTGGCC-3′ 5′-GGCCAGGTCACCACCTGGATGGTGATG-3′	1467-95	459-24	460-11

Underlined letters indicate changed bases. Amino acid residue numbers (minus 19 residues of the signal peptide) and primer location refer to numbering in Hering et al. [32]. Triple mutants were generated by mutating a previous single and double mutant.