Biotechnology Bulletin ›› 2023, Vol. 39 ›› Issue (4): 148-156.doi: 10.13560/j.cnki.biotech.bull.1985.2022-1130
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GUO San-bao1(), SONG Mei-ling2, LI Ling-xin3, YAO Zi-zhao3, GUI Ming-ming2, HUANG Sheng-he1()
Received:
2022-09-15
Online:
2023-04-26
Published:
2023-05-16
GUO San-bao, SONG Mei-ling, LI Ling-xin, YAO Zi-zhao, GUI Ming-ming, HUANG Sheng-he. Cloning and Analysis of Chalcone Synthase Gene and Its Promoter from Euphorbia maculata[J]. Biotechnology Bulletin, 2023, 39(4): 148-156.
引物名称Primer name | 引物序列Primer sequence(5'-3') | 用途Purpose |
---|---|---|
FZL-1 | AAGCAGTGGTATCAACGCAGAGTATTTTTTTTTTTTTTTTTTTTTTTTTVN | cDNA合成 cDNA synthesis |
EmCHS-XF | TGTTCCTGGATGAGGAGATTCTTAC | 3' RACE |
EmCHS-3R | AGCAGTGGTATCAACGCAGAG | |
EmCHS-SR1 | CTTTAAGGTCAGTCATATGCTCGCTCTG | 热不对称交错PCR Tail-PCR |
EmCHS-SR2 | ACGATGGACTCCAGTCCGGCCGCTCTGGGTGATCCTGAAGTAGTAGTC | |
EmCHS-SR3 | ATGGCCTGTAACTCGTGTTGTGCAG | |
ADF1 | ACGATGGACTCCAGAGCGGCCGCVNVNNNGGAA | |
ADF2 | ACGATGGACTCCAGAGCGGCCGCBNBNNNGGTT | |
ADF3 | ACGATGGACTCCAGAGCGGCCGCVVNVNNNCCAA | |
ADF4 | ACGATGGACTCCAGAGCGGCCGCBDNBNNNCGGT | |
ADF0 | ACGATGGACTCCAGAG | |
EmCHS-CF | CCTTAACTCGCTTGTACCTTTGTG | ORF区克隆 ORF clone |
EmCHS-CR | CAGTAGAAAAGTAAAACCACCGATAGG | |
EmCHS-PF | CGGTCCACATGGACAGTAGTG | 启动子克隆 Promoter clone |
EmCHS-PR | CTGTAACTCGTGTTGTGCAGC | |
EmCHS-qF | CGCCGCTAGGGATTAAGGAC | 实时荧光定量PCR RT-qPCR |
EmCHS-qR | CCCGTAATCGCTCAACACCT | |
EmGDI1-qF | TCTAGGTCCCGTTGACGAGT | |
EmGDI1-qR | TCGAGAGCCTTCCCAGTGAT |
Table 1 Primers used in this study
引物名称Primer name | 引物序列Primer sequence(5'-3') | 用途Purpose |
---|---|---|
FZL-1 | AAGCAGTGGTATCAACGCAGAGTATTTTTTTTTTTTTTTTTTTTTTTTTVN | cDNA合成 cDNA synthesis |
EmCHS-XF | TGTTCCTGGATGAGGAGATTCTTAC | 3' RACE |
EmCHS-3R | AGCAGTGGTATCAACGCAGAG | |
EmCHS-SR1 | CTTTAAGGTCAGTCATATGCTCGCTCTG | 热不对称交错PCR Tail-PCR |
EmCHS-SR2 | ACGATGGACTCCAGTCCGGCCGCTCTGGGTGATCCTGAAGTAGTAGTC | |
EmCHS-SR3 | ATGGCCTGTAACTCGTGTTGTGCAG | |
ADF1 | ACGATGGACTCCAGAGCGGCCGCVNVNNNGGAA | |
ADF2 | ACGATGGACTCCAGAGCGGCCGCBNBNNNGGTT | |
ADF3 | ACGATGGACTCCAGAGCGGCCGCVVNVNNNCCAA | |
ADF4 | ACGATGGACTCCAGAGCGGCCGCBDNBNNNCGGT | |
ADF0 | ACGATGGACTCCAGAG | |
EmCHS-CF | CCTTAACTCGCTTGTACCTTTGTG | ORF区克隆 ORF clone |
EmCHS-CR | CAGTAGAAAAGTAAAACCACCGATAGG | |
EmCHS-PF | CGGTCCACATGGACAGTAGTG | 启动子克隆 Promoter clone |
EmCHS-PR | CTGTAACTCGTGTTGTGCAGC | |
EmCHS-qF | CGCCGCTAGGGATTAAGGAC | 实时荧光定量PCR RT-qPCR |
EmCHS-qR | CCCGTAATCGCTCAACACCT | |
EmGDI1-qF | TCTAGGTCCCGTTGACGAGT | |
EmGDI1-qR | TCGAGAGCCTTCCCAGTGAT |
Fig. 2 Cloning of EmCHS gene M: Trans2K® Plus II. 1: PCR product of 3'RACE. 2-3: Tail-PCR product of the second and third rounds. 4: PCR product of ORF
Fig. 7 Expressions of EmCHS from different tissues in different growth stages The different lowercase letters indicate significant differences in EmCHS between different tissues at different growth stages(P <0.05)
元件名称 Element name | 序列 Sequence | 数量 Amount | 功能 Function |
---|---|---|---|
TATA-box | TATA | 6 | -30处基因启动转录核心RNA聚合酶Ⅱ识别位点 Core promoter element around -30 of transcription start |
CAAT-box | CAAT/CAAAT | 11 | 启动子和增强区中常见的顺式作用元件Common cis-acting element in promoter and enhancer region |
MYB | CAACCA | 1 | MYB转录因子结合位点MYB transcription factor binding sites |
MYC | CATTTG | 1 | MYC转录因子结合位点MYC transcription factor binding sites |
Box-II | CCACGTGGC | 1 | 光反应元件的一部分Part of a light responsive element |
Box-IV | ATTAAT | 1 | 参与光反应的保守DNA模块的一部分 Part of a conserved DNA module involved in light responsiveness |
G-box | CACGTG | 1 | 参与光反应的顺式作用调节元件Cis-acting regulatory element involved in light responsiveness |
L-box | ATCCCACCTAC | 1 | 光反应元件的一部分Part of a light responsive element |
ABRE | ACGTG | 3 | 参与脱落酸应答的顺式作用元件Cis-acting element involved in the abscisic acid responsiveness |
ERE | ATTTTAAA | 1 | 乙烯反应元件Ethylene responsive element |
ARE | AAACCA | 1 | 厌氧诱导必须的顺式作用元件Cis-acting regulatory element essential for the anaerobic induction |
STRE | AGGGG | 1 | 应激反应元件Stress responsive element |
WUN-motif | AAATTACT | 1 | 伤口反应元件Wound-responsive element |
ATCT-motif | AATCTAATCC | 1 | 参与光反应的保守DNA模块的一部分 Part of a conserved DNA module involved in light responsiveness |
Table 2 Analysis of EmCHS gene promoter cis-acting elements
元件名称 Element name | 序列 Sequence | 数量 Amount | 功能 Function |
---|---|---|---|
TATA-box | TATA | 6 | -30处基因启动转录核心RNA聚合酶Ⅱ识别位点 Core promoter element around -30 of transcription start |
CAAT-box | CAAT/CAAAT | 11 | 启动子和增强区中常见的顺式作用元件Common cis-acting element in promoter and enhancer region |
MYB | CAACCA | 1 | MYB转录因子结合位点MYB transcription factor binding sites |
MYC | CATTTG | 1 | MYC转录因子结合位点MYC transcription factor binding sites |
Box-II | CCACGTGGC | 1 | 光反应元件的一部分Part of a light responsive element |
Box-IV | ATTAAT | 1 | 参与光反应的保守DNA模块的一部分 Part of a conserved DNA module involved in light responsiveness |
G-box | CACGTG | 1 | 参与光反应的顺式作用调节元件Cis-acting regulatory element involved in light responsiveness |
L-box | ATCCCACCTAC | 1 | 光反应元件的一部分Part of a light responsive element |
ABRE | ACGTG | 3 | 参与脱落酸应答的顺式作用元件Cis-acting element involved in the abscisic acid responsiveness |
ERE | ATTTTAAA | 1 | 乙烯反应元件Ethylene responsive element |
ARE | AAACCA | 1 | 厌氧诱导必须的顺式作用元件Cis-acting regulatory element essential for the anaerobic induction |
STRE | AGGGG | 1 | 应激反应元件Stress responsive element |
WUN-motif | AAATTACT | 1 | 伤口反应元件Wound-responsive element |
ATCT-motif | AATCTAATCC | 1 | 参与光反应的保守DNA模块的一部分 Part of a conserved DNA module involved in light responsiveness |
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