Chinese Bulletin of Botany ›› 2020, Vol. 55 ›› Issue (3): 299-307.DOI: 10.11983/CBB19217
• EXPERIMENTAL COMMUNICATIONS • Previous Articles Next Articles
Huijin Fan1,2,Kangming Jin1,Renying Zhuo1,Guirong Qiao1,*(
)
HomeReceived:2019-11-07
Accepted:2020-03-24
Online:2020-05-01
Published:2020-07-06
Contact:
Guirong Qiao
Huijin Fan, Kangming Jin, Renying Zhuo, Guirong Qiao. Cloning and Expression Analysis of Different Truncated U3 Promoters in Phyllostachys edulis[J]. Chinese Bulletin of Botany, 2020, 55(3): 299-307.
| Primer name | Sequences (5'-3') |
|---|---|
| PeU3-1 | F: ATCCGCCCCTGGTTGCTCTA R: CAAGCCGTTAATCACGCTCTGG |
| PeU3-2 | F: TCAGCGTTGACCTCCTCTTG R: TGGCACGAAATGGGATGAAG |
| 1301-PeU3-1-1-F | F: GACCTGCAGGCATGCAAGCTTTCAGCGTTGACCTCCTCTT R: TTTACCCTCAGATCTACCATTGCTGAGTAGCGCAGGGCTC |
| 1301-PeU3-1-2-F | F: GACCTGCAGGCATGCAAGCTCAGGTCATAAGATGGAAGCT R: TTTACCCTCAGATCTACCATTGCTGAGTAGCGCAGGGCTC |
| 1301-PeU3-1-3-F | F: GACCTGCAGGCATGCAAGCTCAGCCCATACGAAAGTGATT R: TTTACCCTCAGATCTACCATTGCTGAGTAGCGCAGGGCTC |
| 1301-PeU3-2-1-F | F: GACCTGCAGGCATGCAAGCTCCCTAGTGATGCCTTAAAGC R: TTTACCCTCAGATCTACCATTGCATCAATGCTCCAAAGCT |
| 1301-PeU3-2-2-F | F: GACCTGCAGGCATGCAAGCTATGGTATTCTGTTGCGGACT R: TTTACCCTCAGATCTACCATTGCATCAATGCTCCAAAGCT |
| 1301-PeU3-2-3-F | F: GACCTGCAGGCATGCAAGCTTGGCCTTTTTCATGGAGCCG R: TTTACCCTCAGATCTACCATTGCATCAATGCTCCAAAGCT |
| LUC-PeU3-1-1-F | F: TCGACGGTATCGATAAGCTTTTCAGCGTTGACCTCCTCTT R: GCTCTAGAACTAGTGGATCCTGCTGAGTAGCGCAGGGCTC |
| LUC-PeU3-1-2-F | F: TCGACGGTATCGATAAGCTTCAGGTCATAAGATGGAAGCT R: GCTCTAGAACTAGTGGATCCTGCTGAGTAGCGCAGGGCTC |
| LUC-PeU3-1-3-F | F: TCGACGGTATCGATAAGCTTCAGCCCATACGAAAGTGATT R: GCTCTAGAACTAGTGGATCCTGCTGAGTAGCGCAGGGCTC |
| LUC-PeU3-2-1-F | F: TCGACGGTATCGATAAGCTTATCCGCCCCTGGTTGCTCTA R: GCTCTAGAACTAGTGGATCCTGCATCAATGCTCCAAAGCT |
| LUC-PeU3-2-2-F | F: TCGACGGTATCGATAAGCTTATGGTATTCTGTTGCGGACT R: GCTCTAGAACTAGTGGATCCTGCATCAATGCTCCAAAGCT |
| LUC-PeU3-2-3-F | F: TCGACGGTATCGATAAGCTTTGGCCTTTTTCATGGAGCCG R: GCTCTAGAACTAGTGGATCCTGCATCAATGCTCCAAAGCT |
Table 1 Primer sequences used in this study
| Primer name | Sequences (5'-3') |
|---|---|
| PeU3-1 | F: ATCCGCCCCTGGTTGCTCTA R: CAAGCCGTTAATCACGCTCTGG |
| PeU3-2 | F: TCAGCGTTGACCTCCTCTTG R: TGGCACGAAATGGGATGAAG |
| 1301-PeU3-1-1-F | F: GACCTGCAGGCATGCAAGCTTTCAGCGTTGACCTCCTCTT R: TTTACCCTCAGATCTACCATTGCTGAGTAGCGCAGGGCTC |
| 1301-PeU3-1-2-F | F: GACCTGCAGGCATGCAAGCTCAGGTCATAAGATGGAAGCT R: TTTACCCTCAGATCTACCATTGCTGAGTAGCGCAGGGCTC |
| 1301-PeU3-1-3-F | F: GACCTGCAGGCATGCAAGCTCAGCCCATACGAAAGTGATT R: TTTACCCTCAGATCTACCATTGCTGAGTAGCGCAGGGCTC |
| 1301-PeU3-2-1-F | F: GACCTGCAGGCATGCAAGCTCCCTAGTGATGCCTTAAAGC R: TTTACCCTCAGATCTACCATTGCATCAATGCTCCAAAGCT |
| 1301-PeU3-2-2-F | F: GACCTGCAGGCATGCAAGCTATGGTATTCTGTTGCGGACT R: TTTACCCTCAGATCTACCATTGCATCAATGCTCCAAAGCT |
| 1301-PeU3-2-3-F | F: GACCTGCAGGCATGCAAGCTTGGCCTTTTTCATGGAGCCG R: TTTACCCTCAGATCTACCATTGCATCAATGCTCCAAAGCT |
| LUC-PeU3-1-1-F | F: TCGACGGTATCGATAAGCTTTTCAGCGTTGACCTCCTCTT R: GCTCTAGAACTAGTGGATCCTGCTGAGTAGCGCAGGGCTC |
| LUC-PeU3-1-2-F | F: TCGACGGTATCGATAAGCTTCAGGTCATAAGATGGAAGCT R: GCTCTAGAACTAGTGGATCCTGCTGAGTAGCGCAGGGCTC |
| LUC-PeU3-1-3-F | F: TCGACGGTATCGATAAGCTTCAGCCCATACGAAAGTGATT R: GCTCTAGAACTAGTGGATCCTGCTGAGTAGCGCAGGGCTC |
| LUC-PeU3-2-1-F | F: TCGACGGTATCGATAAGCTTATCCGCCCCTGGTTGCTCTA R: GCTCTAGAACTAGTGGATCCTGCATCAATGCTCCAAAGCT |
| LUC-PeU3-2-2-F | F: TCGACGGTATCGATAAGCTTATGGTATTCTGTTGCGGACT R: GCTCTAGAACTAGTGGATCCTGCATCAATGCTCCAAAGCT |
| LUC-PeU3-2-3-F | F: TCGACGGTATCGATAAGCTTTGGCCTTTTTCATGGAGCCG R: GCTCTAGAACTAGTGGATCCTGCATCAATGCTCCAAAGCT |
Figure 1 Construction of expression vectors with GUS/LUC reporter gene driven by different truncated PeU3 promoters
Figure 2 PCR analysis of six truncated PeU3 promoters (A) Amplification using PeU3-1 promoter plasmid as template (1: PeU3-1-3pro; 2: PeU3-1-2pro; 3: PeU3-1-1pro); (B) Amplification using PeU3-2 promoter plasmid as template (1: PeU3-2-3pro; 2: PeU3-2-2pro; 3: PeU3-2-1pro). M: 2 kb DNA marker
Figure 3 Sequence analysis of PeU3 promoters
Figure 4 GUS expression driven by different truncated PeU3 promoters (A) Transient expression of GUS gene driven by different truncated PeU3 promoters in the leaves of Nicotiana benthamiana (Bars=5 mm); (B) Expression of GUS gene driven by different truncated PeU3 promoters in the callus of Dendrocalamus latiflorus (Bars=5 mm). CK: Negative control
Figure 5 LUC expression driven by different truncated PeU3 promoters in the leaves of Nicotiana benthamiana (A), (B) Transient expression LUC assays illustrating the activation of PeU3 promoter in leaves of N. benthamiana; (C) The quantification of the relative luminescence intensities (In each experiment, four biological replications were performed with quanti?cation. Asterisk above the bars denote significant differences determined by the Duncan’s test, P<0.05); CPS: Luminescence intensity
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