Table 1. Site-directed mutagenesis of chagasin.

The pairs of oligonucleotides used to perform site-directed mutagenesis are listed with the respective amino acid substitution. The targeted mutations are marked in bold. T. cruzi Dm28c chagasin gene was used as a template in PCR reactions. The mutations were confirmed by DNA sequencing

Mutation	Construct	Oligonucleotide	Sequence
W93A	pGL1199	OL1695	5′TTTACATGCGCCCGGCGACAGGAGCTTCGC3′
		OL1696	5′GCGAAGCTCCTGTCGCCGGGCGCATGTAAA3′
Y89A	pGL1118	CHAGY1	5′ ATTCTCACTGCCATGCGCCCG 3′
		CHAGY2	5′ CGGGCGCATGGCAGTGAGATT 3′
Y89S	pGL1120	CHAGY3	5′ ATTCTCACTTCCATGCGCCCG 3′
		CHAGY4	5′ CGGGCGCATGGAAGTGAGATT 3′
Y89F	pGL1119	CHAGY5	5′ ATTCTCACTTTCATGCGCCCG 3′
		CHAGY6	5′ CGGGCGCATGAAAGTGAGATT 3′
P30A	pGL1194	OL1679	5′ GCTTCCCAGCAATGCCACCACCGGGTTC 3′
		OL1680	5′ GAACCCGGTGGTGGCATTGCTGGGAAGC 3′
T31A	pGL1209	OL1681	5′ CCAGCAATCCCGCCACCGGGTTCG 3′
		OL1682	5′ CGAACCCGGTGGCGGGATTGCTGG 3′
T31S	pGL1299	OL1851	5′ CCAGCAATCCCTCCACCGGGTTCG 3′
		OL1852	5′ CGAACCCGGTGGAGGGATTGCTGG 3′
T31V	pGL1298	OL1849	5′ CCAGCAATCCCGTCACCGGGTTCG 3′
		OL1850	5′ CGAACCCGGTGACGGGATTGCTGG 3′
T31Y	pGL1210	OL1683	5′ CTTCCCAGCAATCCCTACACCGGGTTCGCGTG3′
		OL1684	5′CACGCGAACCCGGTGTAGGGATTGCTGGGAAG3′
T32A	pGL1211	OL1685	5′ GCAATCCCACCGCCGGGTTCGCGT 3′
		OL1686	5′ ACGCGAACCCGGCGGTGGGATTGC 3′
T32S	pGL1195	OL1687	5′ AGCAATCCCACCTCCGGGTTCGCGTG 3′
		OL1688	5′ CACGCGAACCCGGAGGTGGGATTGCT 3′
T32E	pGL1197	OL1691	5′CCCAGCAATCCCACCGAGGGGTTCGCGTGGTATTTT3′
		OL1692	5′AAAATACCACGCGAACCCCTCGGTGGGATTGCTGGG3′
T32Y	pGL1198	OL1693	5′ CCAGCAATCCCACCTACGGGTTCGCGTGGT 3′
		OL1694	5′ ACCACGCGAACCCGTAGGTGGGATTGCTGG 3′
T32V	pGL1196	OL1689	5′ CAGCAATCCCACCGTCGGGTTCGCGTGG 3′
		OL1690	5′ CCACGCGAACCCGACGGTGGGATTGCTG 3′
T31A/T32A	pGL1258	OL1776	5′ CCCAGCAATCCCGCCGCCGGGTTCGCGTGG 3′
		OL1777	5′ CCACGCGAACCCGGCGGCGGGATTGCTGGG 3′
ΔT31-T32	pGL1259	OL1778	5CAGCTTCCCAGCAATCCCGGGTTCGCGTGGTAT3′
		OL1779	5′ATACCACGCGAACCCGGGATTGCTGGGAAGCTG3′