Acta Horticulturae Sinica ›› 2022, Vol. 49 ›› Issue (9): 1945-1956.doi: 10.16420/j.issn.0513-353x.2021-0794
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JIA Xin1, ZENG Zhen1, CHEN Yue1, FENG Hui2, LÜ Yingmin1,*(), ZHAO Shiwei2,*()
Received:
2022-04-07
Revised:
2022-07-01
Online:
2022-09-25
Published:
2022-10-08
Contact:
LÜ Yingmin,ZHAO Shiwei
E-mail:luyingmin@bjfu.edu.cn;zhaoshiwei@beijingbg.com
CLC Number:
JIA Xin, ZENG Zhen, CHEN Yue, FENG Hui, LÜ Yingmin, ZHAO Shiwei. Cloning and Expression Analysis of RcDREB2A Gene in Rosa chinensis‘Old Blush’[J]. Acta Horticulturae Sinica, 2022, 49(9): 1945-1956.
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URL: https://www.ahs.ac.cn/EN/10.16420/j.issn.0513-353x.2021-0794
用途 Application | 质粒名称 Plasmid name | 引物序列(5′-3′) Primer sequence |
---|---|---|
扩增RcDREB2A全长 Amplification of RcDREB2A full length | RcDREB2A-F | ATGGGTGCTTATGATCAAGGTTCTAATGG |
RcDREB2A-R | GTTTGCATTGCCGCCGCAGCCT | |
荧光定量PCR Quantitative Real-time PCR | RcDREB2A-F1 | GAGCAGGCACATGACTGGATTAGC |
RcDREB2A-R1 | TCTCGGAGCAGGGATGGGTTTAC | |
内参基因荧光定量PCR | RcPP2A-F | GAGGACAGGACCAGGAAGG |
Internal reference genes Quantitative Real-time PCR | RcPP2A-R | GCTCTACGCCGCCAACAT |
亚细胞定位 Subcellular localization | pBI121-RcDREB2A-GFP-F | CATTTACGAACGATACTCGAGATGGGTGCTTATGATCAAGGTTCTAATGG |
pBI121-RcDREB2A-GFP-R | CACCATCACTAGTACGTCGACGTTTGCATTGCCGCCGCAGCCT | |
转录激活活性分析 Transcriptional activation activity analysis | pGBKT7-RcDREB2A-A-F | ATGGAGGCCGAATTCATGGGTGCTTATGATCAAGGTTC |
pGBKT7-RcDREB2A-A-R | CCGCTGCAGGTCGACGGATCCGTTTGCATTGCCGCCGCA | |
pGBKT7-RcDREB2A-N-F | ATGGAGGCCGAATTCATGGGTGCTTATGATCAAGGTTC | |
pGBKT7-RcDREB2A-N-R | CCGCTGCAGGTCGACGGATCCAAAGATTCCTCCTCTGCACAGACC | |
pGBKT7-RcDREB2A-C-F | ATGGAGGCCGAATTCTATAGTGAAAACTGAGGATGGTGAGG | |
pGBKT7-RcDREB2A-C-R | CAGGTCGACGGATCCGTTTGCATTGCCGCCGCA |
Table 1 Related primers
用途 Application | 质粒名称 Plasmid name | 引物序列(5′-3′) Primer sequence |
---|---|---|
扩增RcDREB2A全长 Amplification of RcDREB2A full length | RcDREB2A-F | ATGGGTGCTTATGATCAAGGTTCTAATGG |
RcDREB2A-R | GTTTGCATTGCCGCCGCAGCCT | |
荧光定量PCR Quantitative Real-time PCR | RcDREB2A-F1 | GAGCAGGCACATGACTGGATTAGC |
RcDREB2A-R1 | TCTCGGAGCAGGGATGGGTTTAC | |
内参基因荧光定量PCR | RcPP2A-F | GAGGACAGGACCAGGAAGG |
Internal reference genes Quantitative Real-time PCR | RcPP2A-R | GCTCTACGCCGCCAACAT |
亚细胞定位 Subcellular localization | pBI121-RcDREB2A-GFP-F | CATTTACGAACGATACTCGAGATGGGTGCTTATGATCAAGGTTCTAATGG |
pBI121-RcDREB2A-GFP-R | CACCATCACTAGTACGTCGACGTTTGCATTGCCGCCGCAGCCT | |
转录激活活性分析 Transcriptional activation activity analysis | pGBKT7-RcDREB2A-A-F | ATGGAGGCCGAATTCATGGGTGCTTATGATCAAGGTTC |
pGBKT7-RcDREB2A-A-R | CCGCTGCAGGTCGACGGATCCGTTTGCATTGCCGCCGCA | |
pGBKT7-RcDREB2A-N-F | ATGGAGGCCGAATTCATGGGTGCTTATGATCAAGGTTC | |
pGBKT7-RcDREB2A-N-R | CCGCTGCAGGTCGACGGATCCAAAGATTCCTCCTCTGCACAGACC | |
pGBKT7-RcDREB2A-C-F | ATGGAGGCCGAATTCTATAGTGAAAACTGAGGATGGTGAGG | |
pGBKT7-RcDREB2A-C-R | CAGGTCGACGGATCCGTTTGCATTGCCGCCGCA |
元件名称 Site name | 序列 Sequence | 元件数 Number of sites | 功能 Function |
---|---|---|---|
ABRE | ACGTG | 3 | 脱落酸响应元件Abscisic acid-responsive element |
ATCT-motif | AATCTAATCC | 1 | 光响应元件Light responsive element |
CGTCA-motif | CGTCA | 1 | 茉莉酸响应元件MeJA-responsive element |
MYB | AAACCA | 2 | 干旱诱导相关MYB结合位点MYB binding site involved in drought-inducibility |
G-Box | CACGTT | 2 | 光响应元件Light responsive element |
GARE-motif | TCTGTTG | 1 | 赤霉素响应元件Gibberellin-responsive element |
LTR | CCGAAA | 2 | 低温响应元件Low-temperature responsive element |
P-box | CCTTTTG | 1 | 赤霉素响应元件Gibberellin-responsive element |
TATC-box | TATCCCA | 1 | 赤霉素响应元件Gibberellin-responsive element |
TGA-element | AACGAC | 2 | 生长素响应元件Auxin-responsive element |
TGACG-motif | TGACG | 1 | 茉莉酸响应元件MeJA-responsive element |
Table 2 The cis-acting regulatory elements in the promoter of RcDREB2A
元件名称 Site name | 序列 Sequence | 元件数 Number of sites | 功能 Function |
---|---|---|---|
ABRE | ACGTG | 3 | 脱落酸响应元件Abscisic acid-responsive element |
ATCT-motif | AATCTAATCC | 1 | 光响应元件Light responsive element |
CGTCA-motif | CGTCA | 1 | 茉莉酸响应元件MeJA-responsive element |
MYB | AAACCA | 2 | 干旱诱导相关MYB结合位点MYB binding site involved in drought-inducibility |
G-Box | CACGTT | 2 | 光响应元件Light responsive element |
GARE-motif | TCTGTTG | 1 | 赤霉素响应元件Gibberellin-responsive element |
LTR | CCGAAA | 2 | 低温响应元件Low-temperature responsive element |
P-box | CCTTTTG | 1 | 赤霉素响应元件Gibberellin-responsive element |
TATC-box | TATCCCA | 1 | 赤霉素响应元件Gibberellin-responsive element |
TGA-element | AACGAC | 2 | 生长素响应元件Auxin-responsive element |
TGACG-motif | TGACG | 1 | 茉莉酸响应元件MeJA-responsive element |
Fig. 5 Expression patterns analysis of RcDREB2A in different tissue and under various stress treatments Signifcant differences compared with the control were determined using unpaired Student’s t-tests(**α=0.01).
Fig. 6 Transactivation assay of RcDREB2A The co-transformations of yeast cells with pGBKT7 and pGADT7-T were positive controls,and those with pGBKT7-Lamin and pGADT7-T were negative controls.
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