Table IV.
Gene-specific PCR primers and PCR cycles for semiquantitative RT-PCR amplification
The nucleotide sequences used for designing the PCR primers are shown by their accession number. The designed primer pairs yielded a strong single band for each gene.
| Enzyme/Property | Gene Name | Accession No. | Primer Pair | Amplicon Sizea | No. of Cycles | Synonymous to (Reference) |
|---|---|---|---|---|---|---|
| SS (granule-bound) | GBSSI | AK070431 | F: AACGTGGCTGCTCCTTGAA | 218 | 19 | Waxy (Okagaki, 1992) |
| R: TTGGCAATAAGCCACACACA | ||||||
| SS (soluble) | SSI | AK109458 | F: GGGCCTTCATGGATCAACC | 279 | 22 | |
| R: CCGCTTCAAGCATCCTCATC | ||||||
| SSIIa | AK101978 | F: GGCCAAGTACCAATGGTGAA | 272 | 23 | SSII-3 (Hirose and Terao, 2004; Jiang et al., 2004) | |
| R: GCATGATGCATCTGAAACAAAGC | ||||||
| SSIIIa | AK061604 | F: GCCTGCCCTGGACTACATTG | 334 | 23 | SSIII-2 (Hirose and Terao, 2004; Dian et al., 2005) | |
| R: GCAAACATATGTACACGGTTCTGG | ||||||
| BE | BEI | AK065121 | F: TGGCCATGGAAGAGTTGGC | 191 | 22 | |
| R: CAGAAGCAACTGCTCCACC | ||||||
| BEIIb | AK108535 | F: ATGCTAGAGTTTGACCGC | 261 | 20 | ae (Nishi et al., 2001); RBE3 (Mizuno et al., 1993) | |
| R: AGTGTGATGGATCCTGCC | ||||||
| Debranching enzyme | ISA1 | AB093426 | F: TGCTCAGCTACTCCTCCATCATC | 132 | 22 | Sugary-1 (Kubo et al., 1999) |
| R: AGGACCGCACAACTTCAACATA | ||||||
| PUL | AB012915 | F: ACCTTTCTTCCATGCTGG | 202 | 22 | ||
| R: CAAAGGTCTGAAAGATGGG | ||||||
| α-Amylase | Amy1A | AK101744 | F: GCGCCTGGTGTCAATCAGAA | 309 | 30 | |
| R: CGGATCGGATACAGCTCGTTG | ||||||
| Amy3D | AK119761 | F: TGCACGGCAAGGACTACAGC | 305 | 31 | ||
| R: CCAACGGTTACAAACTGCGTGA | ||||||
| Amy3E | AK064300 | F: GAGCACGCTGGACATCCTCA | 309 | 23 | ||
| R: GCTCGTACACATCTCGCAGCA | ||||||
| AGP (large subunit) | AGPL1 | AK100910 | F: ATGCAGTGCAGTGCGTCTTT | 183 | 23 | OsAPL3 (Akihiro et al., 2005) |
| R: ACTTCACTCGGGGCAGCTTA | ||||||
| AGPL2 | AK071497 | F: CGGGACCGTCATATAAAAGCA | 242 | 19 | OsAPL2 (Akihiro et al., 2005) | |
| R: TCCCATTCCAAAACAAACCA | ||||||
| AGP (small subunit) | AGPS1 | AK073146 | F: AGAATGCTCGTATTGGAGAAAATG | 258 | 22 | OsAPS1 (Akihiro et al., 2005) |
| R: GGCAGCATGGAATAAACCAC | ||||||
| AGPS2b | AK103906 | F: AACAATCGAAGCGCGAGAAA | 186 | 21 | Shrunken (Kawagoe et al., 2005); OsAPS2 (Akihiro et al., 2005) | |
| R: GCCTGTAGTTGGCACCCAGA | ||||||
| SuSy | SuSy2 | AK072074 | F: TTCAGCAGGAGAAGCCGTCAGC | 150 | 22 | RSus2 (Huang et al., 1996) |
| R: CCGGCGTTTATTTGAGGCAAGC | ||||||
| SuSy3 | AK100306 | F: CGGTGAAAAGAATGGGCAATG | 180 | 21 | RSus3 (Huang et al., 1996) | |
| R: CCATGAAAAGGCCAGAGCAT | ||||||
| UDP-Glc pyrophosphorylase | UGP | AK065780 | F: TCCTGGCCCGGTTTAAGTCA | 258 | 23 | |
| R: TGCCGAATGCACACGACAAT | ||||||
| Phospho-Glc isomerase | PGI-a | AK068061 | F: ATCCAGCACATGGCAGCAAA | 324 | 24 | |
| R: AAGGGCACGGGATGACAAGA | ||||||
| PGI-b | AK068236 | F: TGGGGAGTGGAACTGGGAAA | 237 | 26 | ||
| R: CAGAATATGCCGGCTCAACC | ||||||
| Suc transporter | SUT1 | AK100027 | F: AGTTCCGGTCGGTCAGCAT | 241 | 26 | |
| R: ACCGAGGTGGCAACAAAG | ||||||
| GPT | GPT1 | AK060577 | F: AGAAGGGATCCAGATGAAGAATG | 150 | 22 | |
| R: AACAAGAAACGAGCAACATAGACC | ||||||
| ADP-Glc translocator | BT1-2 | AK107368 | F: TGATTGTGCATGGGTGTGATG | 132 | 22 | |
| R: AACAGAGGAAATCGAATCCTACG | ||||||
| PPDK | PPDKA | AJ004965 | F: TTGCAAATGCAGAAACAACCAT | 410 | 26 | |
| R: TGCTGCTAGCCTTGCAATTG | ||||||
| cyPPDKB | D87952 | F: GCTCCGGCTCAATGTGCTCGT | 165 | 24 | ||
| R: CTCCGTCGACACCGTGAAC | ||||||
| chPPDKB | D87952 | F: CCAGGATGCCGTCGGTTTCGA | 368 | NAb | ||
| R: CTCCGTCGACACCGTGAAC | ||||||
| Prolamin | Pro 7 | AF194115 | F: GCGCAGCAGCTACAACTCCA | 206 | 15 | DS16 (this work) |
| R: TGAGCTTATTTTTAACTTCCGAACCA | ||||||
| 13kD Pro | S39468 | F: GGCCATAGCGCACCAGCTAC | 233 | 15 | DS86 (this work) | |
| R: TGTCACATACGATGATACCTGAGC | ||||||
| 16kD Pro | AK107785 | F: TTGCCAGGCTATTTGCACCA | 285 | 15 | ||
| R: CGAACAGCCAAAGACTATTCCAAA | ||||||
| Glutelin | Glu RG55 | AB016501 | F: CCAGCAACAATGCCAACCAG | 207 | 18 | |
| R: TCAGGCCTTGGAGCCTCAAC | ||||||
| Glu type I | AK107285 | F: TTCCGTGCTCTCCCAAATGA | 230 | 14 | DS18 (this work) | |
| R: TGGACAGTACATAGCAGCAAAACA | ||||||
| Glu type II | AK107314 | F: GGTGCATTCACTCCCCTCCA | 225 | 16 | DS72 (this work) | |
| R: CATTGGAACGGGAACACAAAAA | ||||||
| Glu B-1 | AK107343 | F: CGCCGTTCTGGAACGTCAAT | 316 | 14 | DS91 (this work) | |
| R: TTCTTGAGGCTTCGGGCTTG | ||||||
| Globulin | Glb-like | U45322 | F: CGAGAACGGCGAGAAGTGGT | 214 | 21 | |
| R: GCCCTTGCTGAAGCTCGACA | ||||||
| Allergen | RAG2 | AK107328 | F: GTCGACGACAGCTGGTGCAG | 313 | 16 | DS108 (this work) |
| R: TGCTTCCTGACAAATGAAAGCA | ||||||
| PDI | PDI | AK068268 | F: GGATGCAACTGCGAACGATG | 277 | 23 | |
| R: GTCAGGTCCCGTCTCCTCCA | ||||||
| HSP | HSP101 | AK105433 | F: GAGCTGGCCTACAGGGTGGA | 270 | 24 | |
| R: TTCAACGGAGACACCCCACA | ||||||
| HSP82 | AK063751 | F: AGCTCGGCCTCAACATCGAC | 309 | 30 | ||
| R: CGATGGCTTAGAAAGATGTACGC | ||||||
| HSP70 | AK065431 | F: GCCCCAAGATCGAGGAGGTC | 288 | 28 | ||
| R: CAAACAATGCGCACATGCAA | ||||||
| HSP26 | AK063618 | F: AGCGCAGCGTGAGCTCCTAC | 284 | 25 | ||
| R: TCATTCGCTCGTTCGCTGAG | ||||||
| HSP22a | AK063700 | F: CGGTCCTACGGCAGGTTCTG | 315 | 31 | ||
| R: CCGGAACATGTGCTCTGCAA | ||||||
| HSP22b | AK107883 | F: GAGGTTCTGGAGGCGGTTCA | 232 | 34 | ||
| R: GGTGTCGCCGATCACTATTCA | ||||||
| HSP16.9 | AK071240 | F: CGTGCAAGTACCTGCGGATG | 342 | 27 | ||
| R: TCGCATACGGCATACAGACCA | ||||||
| Heat shock transcription factor | HSF | AK106545 | F: TCCAGCTCCAGCCAAACGAT | 255 | 30 | |
| R: CTTGAACTTTCCGCCGCAAC |
a
Numbers indicate the size in bp of amplified fragments.
b
NA, Not amplified.