Multi-locus analysis of human infective Cryptosporidium species and subtypes using ten novel genetic loci

BMC Microbiology

Table 1 List of Cryptosporidium genes selected for this study.

Primer name	Gene function (CryptoDB)	Sequence	Tm (°C)	Annealing temperature (°C)	Size of amplified fragment
cgd2_80 F	ABC transporter family protein	GGA TTG GGG GTG ATA TGT TG	68	60	266 bp
cgd2_80 R		ACC TCC AAG CTG TGT TCC AG	70
cgd6_200 F	Oocyst wall protein 8	CGT TCC AAC AAT GGT GTG TC	68	60	447 bp
cgd6_200 R		GCA GCT GGA GTG CAA TCA TA	68
cgd8_2370 F	Adenosine kinase like ribokinase	CAG GAA TTG CTC ACG GAA AT	66	60	685 bp
cgd8_2370 R		CCT TAA ATG CAT CCC CAC AG	68
Chro.50317 F	RNA polymerase A/beta'/A'' subunit	GAT TTT GAT GGA GGG TCT CG	68	60	752 bp
Chro.50317 R		CTG GCA GCT TCA ACA CCA TA	68
Chro.30149 F	Ubiquitin-protein ligase 1	GGG ATT AGA TGC AGG TGG TG	70	60	331 bp
Chro.30149 R		TGG ATG CTC CAG CAT TAC AT	66
Chro.50457 F	Erythrocyte membrane-associated antigen	CCT TTG GAT TGT CCC GAA TA	66	60	394 bp
Chro.50457 R		CAA TGC CAT ATG ATT TGA GAA AAA	65
cgd6_5020 F	Protein with WD40 repeats	AAC AGG AGC TGA CGA TTG CT	60.4	57	271 bp
cgd6_5020 R		ACA TTG TGC CAT TCC AAG GT	58.35
cgd2_2430 F	Ximpact ortholog conserved protein seen in bacteria and eukaryotes	GTA ACG CAT GGC GAA CCT AT	60.4	57	389 bp
cgd2_2430 R		AAG ATC AGC CTT GCA GCA TT	58.35
Chro.20156 F	Hypothetical protein	TTC GCT TGA AGC CGT AAA CT	58.35	57	247 bp
Chro.20156 R		GGC ATT GAT ACC AGG CAA GT	60.4
Chro.50330 F	Leucyl tRNA synthetase	TCG GTA CAG CAT CAG GTT CA	60.4	57	368 bp
Chro.50330 R		GTT TTT GCT CCC CCA GTT TT	58.35
Cry-15	Oocyst wall protein gene [16]	GTA GAT AAT GGA AGA GAT TGT G	57.08	60	555 bp
Cry-9		GGA CTG AAA TAC AGG CAT TAT CTT G	61.3

Gene name and annotation is according to CryptoDB. For each gene, a set of primers was designed. Primer name is the gene name followed by F or R (for forward and reverse, respectively). For each gene, primer sequences, annealing temperature and PCR product size are detailed.

ISSN: 1471-2180