Biotechnology Bulletin ›› 2023, Vol. 39 ›› Issue (12): 179-186.doi: 10.13560/j.cnki.biotech.bull.1985.2023-0694
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TENG Meng-xin1(), XU Ya1, HE Jing1, WANG Qi1, QIAO Fei2, LI Jing-yang2, LI Xin-guo1()
Received:
2023-07-18
Online:
2023-12-26
Published:
2024-01-11
Contact:
LI Xin-guo
E-mail:tengmx26@163.com;lixinguo13@163.com
TENG Meng-xin, XU Ya, HE Jing, WANG Qi, QIAO Fei, LI Jing-yang, LI Xin-guo. Cloning and Prokaryotic Expression Analysis of MaMC6 in Banana[J]. Biotechnology Bulletin, 2023, 39(12): 179-186.
引物名称 Primer name | 上游引物Forward primer(5'-3') | 下游引物Reverse primer(5'-3') |
---|---|---|
Actin | CGTAGCACCAGAAGAACA | CATAAAGGGAGAGGACAG |
MaMC6 | GTGAGCAACAAGGAACTG | AGTAAAGGCAAGGGTGTT |
pET28a-MaMC6 | AAGAAGGAGATATACATGGAAGCGGGCAAGAAGCG | GTGGTGGTGGTGCTCCATGCAAGCACGTAGCTGCTG |
1302-MaMC6-GFP | CGGGGGACTCTTGACATGGAAGCGGGCAAGAAGCG | TTCTTCTCCTTTACTCATGCAAGCACGTAGCTGCTG |
Table 1 Primers used in the experiment
引物名称 Primer name | 上游引物Forward primer(5'-3') | 下游引物Reverse primer(5'-3') |
---|---|---|
Actin | CGTAGCACCAGAAGAACA | CATAAAGGGAGAGGACAG |
MaMC6 | GTGAGCAACAAGGAACTG | AGTAAAGGCAAGGGTGTT |
pET28a-MaMC6 | AAGAAGGAGATATACATGGAAGCGGGCAAGAAGCG | GTGGTGGTGGTGCTCCATGCAAGCACGTAGCTGCTG |
1302-MaMC6-GFP | CGGGGGACTCTTGACATGGAAGCGGGCAAGAAGCG | TTCTTCTCCTTTACTCATGCAAGCACGTAGCTGCTG |
Fig. 2 Multiple comparison of amino acid sequences between MaMC6 and other plants At: Arabidopsis thaliana; Ma: genome A of Musa acuminata; Mb: genome B of Musa balbisiana; Os: Oryza sativa; Pd: Phoenix dactylifera; Eg: Elaeis guineensis; Ao: Asparagus officinalis; Mi: Mangifera indica; Cs: Citrus sinensis; Zo: Zingiber officinale
Fig. 3 Phylogenetic tree analysis of MaMC6 and other plants Mb: Musa balbisiana; Ma: Musa acuminata; Zo: Zingiber officinale; Os: Oryza sativa; Ao: Asparagus officinalis; Da: Dioscorea alata; Pd: Phoenix dactylifera; Eg: Elaeis guineensis; At: Arabidopsis thaliana; Pv: Pistacia vera; Mi: Mangifera indica; Cs: Citrus sinensis; Tc: Theobroma cacao; Dz: Durio zibethinus; Hs: Hibiscus syriacus; Ga: Gossypium austral; Gh: Gossypium hirsutum; Mc: Momordica charantia; Bh: Benincasa hispida; Ls: Lactuca sativa
Fig. 7 Effects of different calcium effectors on the expression of MaMC6 under salt stress A:100 mmol/L NaCl + 10 mmol/L CaCl2;B:100 mmol/L NaCl + 10 mmol/L EGTA;C:100 mmol/L NaCl + 5 mmol/L LaCl2
Fig. 9 Functional verification of MaMC6 in Escherichia coli BL21 The upper row is the pET28 expressed strain, and the lower row is the pET28a-MaMC6 expressed strain
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