Chinese Journal OF Rice Science ›› 2023, Vol. 37 ›› Issue (6): 577-586.DOI: 10.16819/j.1001-7216.2023.230108
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QI Panpan1,2, GUO Liuming2, LI Jing2, LÜ Mingfang2, YUAN Zhengjie2, ZHANG Hengmu1,2,*()
Received:
2023-01-28
Revised:
2023-03-10
Online:
2023-11-10
Published:
2023-11-14
Contact:
*email: zhhengmu@tsinghua.org.cn
齐盼盼1,2, 郭留明2, 李静2, 吕明芳2, 袁正杰2, 张恒木1,2,*()
通讯作者:
*email: zhhengmu@tsinghua.org.cn
基金资助:
QI Panpan, GUO Liuming, LI Jing, LÜ Mingfang, YUAN Zhengjie, ZHANG Hengmu. cDNA Cloning and Molecular Characterization of OsTAF12b Gene in Oryza sativa[J]. Chinese Journal OF Rice Science, 2023, 37(6): 577-586.
齐盼盼, 郭留明, 李静, 吕明芳, 袁正杰, 张恒木. 水稻TAF12b基因cDNA克隆及其分子特性鉴定[J]. 中国水稻科学, 2023, 37(6): 577-586.
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URL: http://www.ricesci.cn/EN/10.16819/j.1001-7216.2023.230108
引物名称 Primer name | 引物序列 Sequence (5'-3') | 注释 Annotation |
---|---|---|
5'R1 | GCCTGCTGCTGTTTCTGAGGAAGGG | 5'RACE |
5'R2 | GATTACGCCAAGCTTTGTGGAGACAGTGTGCCTT | |
3'R1 | CAGCCAAGGCAGAAGGGTATGGTGC | 3'RACE |
3'R2 | GATTACGCCAAGCTTGGGTGGTATGAGAGGAAATG | |
OsTAF12b-F | TCAAACTACCCCACAGGACC | OsTAF12b亚细胞定位载体构建 Construction of subcellular localization vector of OsTAF12b |
OsTAF12b-R | GCGAACAATCTGTCAGTCAG | |
pCV-OsTAF12b-GFP-F | ACTCTAGACCCCTGGGATCCATGGCGGATCCGCCGTCCG | |
pCV-OsTAF12b-GFP-R | GCCCTTGCTCACCATGTCGACGAAACGGGGCACTTTTGAC | |
pCV-GFP-OsTAF12b-F | GACGAGCTGTACAAGGTCGACATGGCGGATCCGCCGTCCG | |
pCV-GFP-OsTAF12b-R | TTCGAGCTCGCCTGGGGATCCTTAGAAACGG GGCACTTTT | |
pCV-H2B-mCherry-F | ACTCTAGACCCCTGGGATCCATGGCGAAGGCAGATAAGA | 核定位标记载体构建Construction of nuclear localization marker vector |
pCV-H2B-mCherry-R | GCCCTTGCTCACCATGTCGACAGCAGAACTCGTAAACTTC | |
qActin-F | GTATCCATGAGACTACATACAACT | 内参基因定量引物 Primers of internal reference gene |
qActin-R | TACTCAGCCTTGGCAATCCACA | |
qOs12b-F | GCATCCTCATCTCAATACCC | OsTAF12b定量引物 qRT-PCR primers of OsTAF12b |
qOs12b-R | CCAGTCATAGTTCCACCTGAT |
Table 1. Information of primers used in the study.
引物名称 Primer name | 引物序列 Sequence (5'-3') | 注释 Annotation |
---|---|---|
5'R1 | GCCTGCTGCTGTTTCTGAGGAAGGG | 5'RACE |
5'R2 | GATTACGCCAAGCTTTGTGGAGACAGTGTGCCTT | |
3'R1 | CAGCCAAGGCAGAAGGGTATGGTGC | 3'RACE |
3'R2 | GATTACGCCAAGCTTGGGTGGTATGAGAGGAAATG | |
OsTAF12b-F | TCAAACTACCCCACAGGACC | OsTAF12b亚细胞定位载体构建 Construction of subcellular localization vector of OsTAF12b |
OsTAF12b-R | GCGAACAATCTGTCAGTCAG | |
pCV-OsTAF12b-GFP-F | ACTCTAGACCCCTGGGATCCATGGCGGATCCGCCGTCCG | |
pCV-OsTAF12b-GFP-R | GCCCTTGCTCACCATGTCGACGAAACGGGGCACTTTTGAC | |
pCV-GFP-OsTAF12b-F | GACGAGCTGTACAAGGTCGACATGGCGGATCCGCCGTCCG | |
pCV-GFP-OsTAF12b-R | TTCGAGCTCGCCTGGGGATCCTTAGAAACGG GGCACTTTT | |
pCV-H2B-mCherry-F | ACTCTAGACCCCTGGGATCCATGGCGAAGGCAGATAAGA | 核定位标记载体构建Construction of nuclear localization marker vector |
pCV-H2B-mCherry-R | GCCCTTGCTCACCATGTCGACAGCAGAACTCGTAAACTTC | |
qActin-F | GTATCCATGAGACTACATACAACT | 内参基因定量引物 Primers of internal reference gene |
qActin-R | TACTCAGCCTTGGCAATCCACA | |
qOs12b-F | GCATCCTCATCTCAATACCC | OsTAF12b定量引物 qRT-PCR primers of OsTAF12b |
qOs12b-R | CCAGTCATAGTTCCACCTGAT |
Fig. 1. Amplification of OsTAF12b cDNA. M, DNA marker; Line 1, Amplified fragment of central region; Line 2, 5'-RACE product; Line 3, 3'-RACE product.
Fig. 2. Schematic graph of OsTAF12b gene structure. A, Location of OsTAF12b gene on rice Chromosome 1; B, Schematic graph of four transcripts of OsTAF12b. The white boxes represent the untranslated regions; the gray boxes represent the coding regions; the black lines represent the introns; the dotted lines indicate the alternative splicing sites.
Fig. 5. Subcellular localization of OsTAF12b in Nicotiana benthamiana leaf epidermal cells(A) and rice protoplasts(B). A, Localization of OsTAF12b protein in Nicotiana benthamiana leaf epidermal cells(scale bar 50 μm); B, Localization of OsTAF12b protein in rice protoplasts(scale bar 5 μm).
Fig. 6. Expression patterns of OsTAF12b. A, Relative expression level of OsTAF12b in different tissues of rice; B, Relative expression level of OsTAF12b under heat treatment; C, Relative expression level of OsTAF12b under cold treatment; D and F, Relative expression level of OsTAF12b under PEG treatment; E and G, Relative expression level of OsTAF12b under NaCl treatment. Data are mean±SD from three independent experiments. The asterisks on the top of the columns indicate significant differences from the value at 0 h (ns, not significant; *P < 0.05; **P < 0.01).
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