Biotechnology Bulletin ›› 2023, Vol. 39 ›› Issue (11): 261-269.doi: 10.13560/j.cnki.biotech.bull.1985.2023-0133
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YANG Xu-yan(), ZHAO Shuang, MA Tian-yi, BAI Yu, WANG Yu-shu()
Received:
2023-02-15
Online:
2023-11-26
Published:
2023-12-20
Contact:
WANG Yu-shu
E-mail:yangxy1219@126.com;wangys1019@126.com
YANG Xu-yan, ZHAO Shuang, MA Tian-yi, BAI Yu, WANG Yu-shu. Cloning of Three Cabbage WRKY Genes and Their Expressions in Response to Abiotic Stress[J]. Biotechnology Bulletin, 2023, 39(11): 261-269.
应用Application | 引物名称Primer name | 引物序列Primer sequence(5'-3') |
---|---|---|
基因克隆Gene cloning | BoWRKY40 | F: ATGGACCAGTACCCATCGTCTTTG R: CTACTTGTCGGTTTGATTCTGTTGG |
BoWRKY46 | F: ATGTTAATGGAAGAAAAGCTTGTG R: TTACATCCACGACAAATCTTGAG | |
BoWRKY70 | F: ATGGATATTGTTAGTAATAACAAAGC R: TCATTGTCGTTGAACCGGAAATC | |
实时荧光定量PCR Real-time PCR | qBoWRKY40 | F: GGTGCTGCCAACAGAAGTAGGAG R: GGGCTTGTCACTTTCTTGGTTTCAG |
qBoWRKY46 | F: AAGAGGTTGCTTGGCGGTGAAG R: AGAGTGTTTGGTGGTTGATGGAGTG | |
qBoWRKY70 | F: AACCCATCCCCTCCTATCTCTTCAC R: ATCCTCAAGTTTGCCGTCGTTGG | |
BoActin2 | F: CGTGACCTTACTGACTACC R: CTCCATCTCCTGCTCGTA | |
表达载体Expression vector | BoWRKY40 | F: ctgcaggggcccggggtcgacATGGACCAGTACCCATCGTCTTTG R: gcccttgctcaccatggtaccCTACTTGTCGGTTTGATTCTGTTGG |
BoWRKY46 | F: ctgcaggggcccggggtcgaccATGTTAATGGAAGAAAAGCTTGTG R: gcccttgctcaccatggtaccTTACATCCACGACAAATCTTGAG | |
BoWRKY70 | F:ctgcaggggcccggggtcgacATGGATATTGTTAGTAATAACAAAGC R: gcccttgctcaccatggtaccTCATTGTCGTTGAACCGGAAATC |
Table 1 List of primers
应用Application | 引物名称Primer name | 引物序列Primer sequence(5'-3') |
---|---|---|
基因克隆Gene cloning | BoWRKY40 | F: ATGGACCAGTACCCATCGTCTTTG R: CTACTTGTCGGTTTGATTCTGTTGG |
BoWRKY46 | F: ATGTTAATGGAAGAAAAGCTTGTG R: TTACATCCACGACAAATCTTGAG | |
BoWRKY70 | F: ATGGATATTGTTAGTAATAACAAAGC R: TCATTGTCGTTGAACCGGAAATC | |
实时荧光定量PCR Real-time PCR | qBoWRKY40 | F: GGTGCTGCCAACAGAAGTAGGAG R: GGGCTTGTCACTTTCTTGGTTTCAG |
qBoWRKY46 | F: AAGAGGTTGCTTGGCGGTGAAG R: AGAGTGTTTGGTGGTTGATGGAGTG | |
qBoWRKY70 | F: AACCCATCCCCTCCTATCTCTTCAC R: ATCCTCAAGTTTGCCGTCGTTGG | |
BoActin2 | F: CGTGACCTTACTGACTACC R: CTCCATCTCCTGCTCGTA | |
表达载体Expression vector | BoWRKY40 | F: ctgcaggggcccggggtcgacATGGACCAGTACCCATCGTCTTTG R: gcccttgctcaccatggtaccCTACTTGTCGGTTTGATTCTGTTGG |
BoWRKY46 | F: ctgcaggggcccggggtcgaccATGTTAATGGAAGAAAAGCTTGTG R: gcccttgctcaccatggtaccTTACATCCACGACAAATCTTGAG | |
BoWRKY70 | F:ctgcaggggcccggggtcgacATGGATATTGTTAGTAATAACAAAGC R: gcccttgctcaccatggtaccTCATTGTCGTTGAACCGGAAATC |
Fig. 2 Cis-elements of three BoWRKY genes ABRE: Abscisic acid responsiveness; ARE: anaerobic induction; AuxRR-core: auxin responsiveness; CAT-box: meristem expression; CGTCA-motif: MeJA-responsiveness; G-box: light responsiveness; MBS: MYB binding site involved in drought-inducibility; TCA-element: salicylic acid responsiveness; GARE-motif: gibberellin-responsive; LTR: low-temperature responsiveness; TC-rich repeats: defense and stress responsiveness; W-box: WRKY binding site
Fig. 3 Multiple alignment of conserved domains of 3 BoWRKYs and WRKY from other plant species Bo: B. oleracea var. capitata; Bra: Brassica napus; At: Arabidopsis thaliana; Gh: Gossypium hirsutum. The same below
Fig. 7 Expression patterns of BoWRKYs gene under abiotic stresses The error line in the figure refers to the standard deviation. The lowercase letters indicate that the gene expression level reached(P<0.05)significant level at different treatment times
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