Biotechnology Bulletin ›› 2023, Vol. 39 ›› Issue (5): 217-223.doi: 10.13560/j.cnki.biotech.bull.1985.2022-1231
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JIANG Qing-chun(), DU Jie, WANG Jia-cheng, YU Zhi-he, WANG Yun, LIU Zhong-yu()
Received:
2022-10-08
Online:
2023-05-26
Published:
2023-06-08
Contact:
LIU Zhong-yu
E-mail:2870140725@qq.com;zyliu2004@126.com
JIANG Qing-chun, DU Jie, WANG Jia-cheng, YU Zhi-he, WANG Yun, LIU Zhong-yu. Expression and Function Analysis of Transcription Factor PcMYB2 from Polygonum cuspidatum[J]. Biotechnology Bulletin, 2023, 39(5): 217-223.
引物名称 Primer name | 引物序列 Primer sequence(5'-3') | 退火温度 Annealing temperature/℃ |
---|---|---|
35Sf3 | CGGGATCCCATGGAGTCAAAGATTCA | 55 |
35Sr3 PcMYB2-F PcMYB2-R | AAAACTGCAGAGTCCCCCGTGTTCTCTC CGCGGATCCGTATGGGGAGAAGAAA AAAACTGCAGGAAATTTCCGAGCCATCC | 55 |
qPcActin-F1 | GTCCCTGCCATGTATGTTGC | 58 |
qPcActin-R1 | ACCATCACCAGAATCCAGCA | |
qPcMYB2-F | GAGGAAGGGTGGAGAAATGA | 58 |
qPcMYB2-R | TGCCCTTGTGCTGTATCTGA |
Table 1 Primer sequence information
引物名称 Primer name | 引物序列 Primer sequence(5'-3') | 退火温度 Annealing temperature/℃ |
---|---|---|
35Sf3 | CGGGATCCCATGGAGTCAAAGATTCA | 55 |
35Sr3 PcMYB2-F PcMYB2-R | AAAACTGCAGAGTCCCCCGTGTTCTCTC CGCGGATCCGTATGGGGAGAAGAAA AAAACTGCAGGAAATTTCCGAGCCATCC | 55 |
qPcActin-F1 | GTCCCTGCCATGTATGTTGC | 58 |
qPcActin-R1 | ACCATCACCAGAATCCAGCA | |
qPcMYB2-F | GAGGAAGGGTGGAGAAATGA | 58 |
qPcMYB2-R | TGCCCTTGTGCTGTATCTGA |
Fig. 2 Structure of pCAMBIA 1380-35S-PcMYB2 vectors LB: Left border of transfer DNA. t35S: Cauliflower mosaic virus CaMV 35S terminator. hpt: Hygromycin resistance gene P35S - Cauliflower mosaic virus CaMV 3SS promoter. tNOS: Nitric oxide synthase gene terminator. RB: Right of transfer DNA boundary
Fig. 5 Expression pattern analysis of PcMYB2 A : Expressions of PcMYB2 gene in different tissues. B : PcMYB2 expressions in leaves under exogenous signaling molecules. ** P < 0.01. The same below
Fig. 6 Verification of recombinant plamid pCAMBIA1380-35S-PcMYB2 M: Marker. 1: Digestion recombinant plasmid by BamH I/Pst I. 2: Digestion recombinant plasmid by BamH I/ Nco I
Fig. 7 Identification and phenotypic observation of trans-genic lines A: RT-qPCR identification. B: Observation on the seed coat's color of A. thaliana. C: Hairy roots before DMACA staining. D: Hairy root after DMACA staining. CK1: Wild-type A.thaliana; A1: transgenic A.thaliana; CK2: wild-type hairy roots; H1: transgenic hairy roots
Fig. 8 Determination of proanthocyanidins CK1: Wild-type A.thaliana; A1: transgenic A. thaliana; CK2: wild-type hairy roots; H1: transgenic hairy roots; *P < 0.05
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