The tandemly repeated NTPase (NTPDase) from Neospora caninum is a canonical dense granule protein whose RNA expression, protein secretion and phosphorylation coincides with the tachyzoite egress

Table 1 Primers used to amplify NcNTPase, NcGRA7, NcSAG1 and NcTUBα sequences by real time-PCR

Protein	ToxoDB accession number	Primer sequences	Reference	Length	Introns ^a	Slope ^b	R^{2 b}
NcNTPase	NCLIV_068400	Fw-ATTGACCCCGACAGTATTCG Rv-ACGCTTGGAATCAACAGACCT	This study	129 bp; Pos. 283–411	No	-3.54	0.998
NcGRA7	NCLIV_021640	Fw-GAACAGCATGAAGGGGACAT Rv-CACCATCTGTAATGGCATCG	This study	130 bp; Pos. 97–226	No	-3.61	0.997
NcSAG1	NCLIV_033230	Fw-CGGTGTCGCAATGTGCTCTT Rv-ACGGTCGTCCCAGAACAAAC	[70]	150 bp; Pos. 504–653	No	-3.24	0.997
NcTUBα	NCLIV_058890	Fw-GGTAACGCCTGCTGGGAG Rv-GCTCCAAATCCAAGAAGACGCA	[71]	166 bp; Pos. 49–214	Yes*	-3.24	0.994

^aPrimers for intron-containing sequences were designed using cDNA as template. * Forward primer for NcTUBα amplification annealed at intron splice junction to prevent amplification of genomic DNA
^bDescriptive values of real time-PCR from standard curves for each pair of primers are shown

ISSN: 1756-3305