Table 1 Specific primer pairs used for ORF cloning, qPCR, and recombinant plasmid construction. 26S-2 was used as an internal control genes
Genes | Primers(5′-3′) | Usage |
|---|---|---|
PnPht1;1 | F: ATGTCTGGGAATAATCTGCAGG | to amplify cDNA ORF |
R: CTAAACTTGTGCCTCATTTGCA | ||
PnPht1;2 | F: ATGGCTCGAGAGCAACTAGAAG | |
R: TTATACCGGCACAGTCCTGTTAG | ||
qPnPht1;1 | F:GAACGGGAATTTTGGTTGCTG | to amplify segments for qPCR |
R:AAGTCGCCTTGTGGCTGAGTG | ||
qPnPht1;2 | F:GCGCGTTTATAGCGGCTGTTT | |
R:ACTTTTGCCTCTTGGGGGTCA | ||
26S-2(reference gene) | F:CAGTATTTAGCCTTGGACGGAATT | |
R:CGGGTTGTTTGGGAATGC | ||
SPnPht1;1(BamH I/Kpn I) | F: CGCGGATCCATGTCTGGGAATAATCTGCAGG | to amplify genes for recombinant plasmid construction in S. cerevisiae |
R: CGGGGTACCCTAAACTTGTGCCTCATTTGCA | ||
SPnPht1;2(Xba I/ Xma I) | F: TGCTCTAGAATGGCTCGAGAGCAACTAGAAG | |
R:TCCCCCCGGGTTATACCGGCACAGTCCTGTTAG | ||
NbPnPht1;1(BamH I/Sal I) pENTR | F:GAGAACACGGGGGACTGGTACCCGGGGATCCATGTCTGGGAATAATCTGCAGG | to amplify genes for recombinant plasmid construction in N. benthamiana |
R:ACAGCTCCTCGCCCTTGCTCACCATGTCGACAACTTGTGCCTCATTTGCATCCT | ||
NbPnPht1;2(BamH I/Sal I) pENTR | F:GAGAACACGGGGGACTGGTACCCGGGGATCCATGGCTCGAGAGCAACTA | |
R:ACAGCTCCTCGCCCTTGCTCACCATGTCGAC ACGAATGGCCCTTTTATCC | ||
NbUPF0057(BamH I/Sal I) pENTR | F:GAGAACACGGGGGACTGGTACCCGGGGATCCATGGTCTCAAGATGTGCA | |
R:ACAGCTCCTCGCCCTTGCTCACCATGTCGACAGCAAGAGTGTCATAACG |