Biotechnology Bulletin ›› 2022, Vol. 38 ›› Issue (10): 184-194.doi: 10.13560/j.cnki.biotech.bull.1985.2021-1546
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LUO Ying(), TANG Zhi, WANG Fan, LIU Xiao-xia, LUO Xiao-fang, HE Fu-lin()
Received:
2021-12-14
Online:
2022-10-26
Published:
2022-11-11
Contact:
HE Fu-lin
E-mail:yingluo_301@163.com;2339695475@qq.com
LUO Ying, TANG Zhi, WANG Fan, LIU Xiao-xia, LUO Xiao-fang, HE Fu-lin. Cloning and Response Analysis to Abiotic Stress of GbR2R3-MYB1 Gene from Ginkgo biloba[J]. Biotechnology Bulletin, 2022, 38(10): 184-194.
引物名称Primer name | 引物序列Primer sequence(5'-3') | 用途Purpose |
---|---|---|
GbR2R3MYB1-F | AGAAGCTGCTCCTCGTTACAG | 目的基因扩增引物 |
GbR2R3MYB1-R | GTCCTTGATCAAAATGGGGTA | 目的基因扩增引物 |
pC007-F | TGTAAAACGACGGCCAGT | pClone007载体引物 |
pC007-R | CAGGAA ACAGCTATGACC | pClone007载体引物 |
GFP-F | AGAACACGGGGGACGAGCTCGGTACCATGGGTCGGTCTCCTTGCTG | 亚细胞定位引物 |
GFP-R | CCCTTGCTCACCATGTCGACTCTAGATACCCGCAGTTGCCTGTAAT | 亚细胞定位引物 |
Gb18S-F | CGAAGACGATCAGATACCG | qPCR内参基因 |
Gb18S-R | TCAGCCTTGCGACCATAC | qPCR内参基因 |
q-R2R3MYB1-F | ACGGCAGCTCCAGCTAATTC | qPCR检测引物 |
q-R2R3MYB1-R | GTGGTGGCTCTGCTTATGCT | qPCR检测引物 |
Table 1 Primer sequences used in this study
引物名称Primer name | 引物序列Primer sequence(5'-3') | 用途Purpose |
---|---|---|
GbR2R3MYB1-F | AGAAGCTGCTCCTCGTTACAG | 目的基因扩增引物 |
GbR2R3MYB1-R | GTCCTTGATCAAAATGGGGTA | 目的基因扩增引物 |
pC007-F | TGTAAAACGACGGCCAGT | pClone007载体引物 |
pC007-R | CAGGAA ACAGCTATGACC | pClone007载体引物 |
GFP-F | AGAACACGGGGGACGAGCTCGGTACCATGGGTCGGTCTCCTTGCTG | 亚细胞定位引物 |
GFP-R | CCCTTGCTCACCATGTCGACTCTAGATACCCGCAGTTGCCTGTAAT | 亚细胞定位引物 |
Gb18S-F | CGAAGACGATCAGATACCG | qPCR内参基因 |
Gb18S-R | TCAGCCTTGCGACCATAC | qPCR内参基因 |
q-R2R3MYB1-F | ACGGCAGCTCCAGCTAATTC | qPCR检测引物 |
q-R2R3MYB1-R | GTGGTGGCTCTGCTTATGCT | qPCR检测引物 |
Fig.1 Agarose gel electrophoresis of PCR amplified prod-ucts of GbR2R3-MYB1 A:PCR amplification of target fragment of GbR2R3-MYB1. B:Bacteria liquid PCR;M:DL2000 DNA marker;1-3:positive clones of colony PCR identification
Fig. 7 Prediction of secondary structure for GbR2R3-MYB1 protein Blue:Alpha helix. Green:Beta turn. Purple:Random coil. Red:Extended strand. D:Tertiary structure analysis of GbR2R3-MYB1 protein
Fig.10 Expression analysis of GbR2R3-MYB1 gene under salt,drought,cold,and heat stresses Each value was calculated as the mean of three samples standard error(SE). The different letters above the bar indicate significant difference(P < 0.05)and very significant difference(P < 0.01)
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