Gene expression and proteomic analysis of the formation of Phakopsora pachyrhizi appressoria

Stone, Christine L; McMahon, Michael B; Fortis, Laurie L; Nuñez, Alberto; Smythers, Gary W; Luster, Douglas G; Frederick, Reid D

doi:10.1186/1471-2164-13-269

BMC Genomics

Table 1 Primer pairs used for real-time quantitative reverse transcription PCR analysis of expression patterns of Phakopsora pachyrhizi ESTs

From: Gene expression and proteomic analysis of the formation of Phakopsora pachyrhizi appressoria

Clone/contig	Target gene product		Primer sequences	Amplicon size (bp)	Primer (nM)	Annealing temperature (°C)
Pp3004¹	serine/threonine-protein kinase	Forward	GGAACCAACGTCGACAAGAG	130	200	60
		Reverse	CTGGTCCTCCTCAACCTCAG
Pp3243¹	prefoldin subunit 5	Forward	AACCTGATCAACTGGCATCC	81	200	59
		Reverse	CCTTTAGTTGCCCAAACGAG
Pp3282¹	ubiquitin-protein ligase	Forward	AAATTCCTGGAGACGATTGC	150	300	60
		Reverse	TTGCACTACCTTGTGCGGTA
Pp3495¹	G2/M phase checkpoint control protein Sum2	Forward	CAGCCTAGAACAGGTCAGATCC	105	200	58
		Reverse	GCCCGGAAAACGATAAATTC
Pp3505¹	hypothetical protein	Forward	GAGACAAGCCCCATTGAGAG	104	200	60
		Reverse	GTCTTTGGCAGGGTCTTCTG
Pp3684¹	HMG-CoA reductase	Forward	CGACAGCTTGCTCGAATTATC	87	200	60
		Reverse	CTTAACCAAGTGTCCAGCTGC
contig2F¹	5-aminolevulinate synthase	Forward	GATGAGGTTCACGCCATTG	117	200	60
		Reverse	GTCCACCCGATCCATTACAC
Pp3186¹	hypothetical protein	Forward	AAGGACTGGTGAGGTTGGTG	76	200	62
		Reverse	TGCTCTACCACCGTAGGACC
contig2N¹	P-type cation-transporting ATPase	Forward	GCCTGAAGTGATTCCTCAGC	124	400	55
		Reverse	ATCCCAACAAACGAAAGTG
Pp3042²	GTPase-binding protein	Forward	TGCCCAGAAAATTGGTTCTC	102	300	58
		Reverse	GCCAAAAGTGCATACCGAGT
Pp3205²	NADPH oxidase A	Forward	ACCCAAGGCTGCCATTAGTA	134	300	56
		Reverse	GGTGCCCTCCATAAAACAAA
Pp3222²	septin/cell division control/GTP binding protein	Forward	TCCTCCCAAAGAACATCCAG	135	300	60
		Reverse	AAGTCTCCATAGCCCGGAGT
Pp3409²	MFS sugar transporter	Forward	CGGATGTAGCATGGAGACTAATG	123	300	60
		Reverse	CATCTGAAATCCACCCCTTAGA
Pp3717²	tripeptidyl-peptidase 1 precursor	Forward	GGCGGAGGGTTTTCAAATTA	116	300	59
		Reverse	CTTCCTGACCCATCGAACAT
Pp3888²	microtubule associated protein	Forward	GGGATCAGCTCTACGTCTGC	150	300	60
		Reverse	AGATTGCCAGCAGCCTTTTA
Pp3944²	autophagy-related protein 8	Forward	CCGTATTCCTGTCATCTGTGAG	107	300	60
		Reverse	CATAGACGAACTGCCCAACC
Pp3998²	delta (12) fatty acid desaturase	Forward	CCCTCTCCTCCCTCACTACC	135	300	59
		Reverse	TTGGAGCACAGATGATGAGC
Pp4000²	subtilase-type proteinase psp3	Forward	TCACTTTGACATTGGAACGAG	93	300	60
		Reverse	GCAACCTCAGGAAGGGTTCT
contig2S²	acyl-CoA dehydrogenase	Forward	AATCGATACCGCCGTCTATG	124	300	60
		Reverse	CCTGTGTAAGCCGAAGAAGG
contig6B²	conidiation-related protein	Forward	AATGACAGAGGTGGCGAGAC	107	300	60
		Reverse	TGTCGAAGCTCGTCCTTTTT
Pp481²	MAS3 protein	Forward	CGTGATGGTACTCGAACGAAC	68	200	62
		Reverse	CTTGAAACCTCACGGTCTCG
NA³	P. pachyrhizi	Forward	CCAAGGCTTCTTCGTGTTTCA	65	200	60
	α-tubulin	Reverse	CAAGAGAAGAGCGCCAAACC

¹Primer pairs used for quantification of mRNA transcripts throughout the infection cycle.
²Primer pairs used for quantification of mRNA transcripts during germination, germ tube elongation, and appressoria formation.
³Primer pairs used for both quantitative analyses.

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ISSN: 1471-2164

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