Biotechnology Bulletin ›› 2021, Vol. 37 ›› Issue (8): 121-130.doi: 10.13560/j.cnki.biotech.bull.1985.2020-1516
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FAN Ya-peng(), RUI Cun, ZHANG Yue-xin, CHEN Xiu-gui, LU Xu-ke, WANG Shuai, ZHANG Hong, XU Nan, WANG Jing, CHEN Chao, YE Wu-wei()
Received:
2020-12-15
Online:
2021-08-26
Published:
2021-09-10
Contact:
YE Wu-wei
E-mail:fanyp0605@126.com;yew158@163.com
FAN Ya-peng, RUI Cun, ZHANG Yue-xin, CHEN Xiu-gui, LU Xu-ke, WANG Shuai, ZHANG Hong, XU Nan, WANG Jing, CHEN Chao, YE Wu-wei. Cloning,Expression and Preliminary Bioinformatics Analysis of the Alkaline Tolerant Gene GhZAT12 in Gossypium hirsutum[J]. Biotechnology Bulletin, 2021, 37(8): 121-130.
引物名称Primer name | 引物序列Primers sequence(5'-3') | 描述Description |
---|---|---|
GhZAT12 | F:ATGAAGAGAGGCAGAGATATTGATG | GhZAT12基因引物 Primer pairs for GhZAT12 gene |
R:CTATATAAAACAATGAACAACCGGA | ||
GFP-GhZAT12 | F:AGAACACGGGGGACTCTAGAATGAAGAGAGGCAGAGATATTG | GhZAT12亚细胞定位载体引物 Primer pairs for subcellular localization vector |
R:TAGTCAGGCGCGCCGGTACCTATAAAACAATGAACAACCGG | ||
Actin(AY305733) | F:ATCCTCCGTCTTGACCTTG | 内参基因引物 Primer pairs for internal reference gene |
R:TGTCCGTCAGGCAACTCAT | ||
QGhZAT12 | F:CTGCTCTGAATGACGGGTTG | GhZAT12荧光定量引物 Primer pairs for qRT-PCR of GhZAT12 |
R:CCGGAGTTGGTGTCACTTTC |
Table 1 Primer pairs used in the experiments
引物名称Primer name | 引物序列Primers sequence(5'-3') | 描述Description |
---|---|---|
GhZAT12 | F:ATGAAGAGAGGCAGAGATATTGATG | GhZAT12基因引物 Primer pairs for GhZAT12 gene |
R:CTATATAAAACAATGAACAACCGGA | ||
GFP-GhZAT12 | F:AGAACACGGGGGACTCTAGAATGAAGAGAGGCAGAGATATTG | GhZAT12亚细胞定位载体引物 Primer pairs for subcellular localization vector |
R:TAGTCAGGCGCGCCGGTACCTATAAAACAATGAACAACCGG | ||
Actin(AY305733) | F:ATCCTCCGTCTTGACCTTG | 内参基因引物 Primer pairs for internal reference gene |
R:TGTCCGTCAGGCAACTCAT | ||
QGhZAT12 | F:CTGCTCTGAATGACGGGTTG | GhZAT12荧光定量引物 Primer pairs for qRT-PCR of GhZAT12 |
R:CCGGAGTTGGTGTCACTTTC |
Fig.2 Multiple sequence alignment of homologous amino acids in GhZAT12 protein XP_002314198.3 Populus trichocarpa, XP_002284111.1 Vitis vinifera, XP_017982131.1 Theobroma cacao, XP_008654280.1 Zea mays, NP_200790.1 Arabidopsis thaliana, NP_001237020.2 Glycine max, XP_021628881.1 Manihot esculenta, XP_002533110.1 Ricinus communis, XP_021887642.1 Carica papaya, XP_004147699.1 Cucumis sativus, XP_006345939.1 Solanum tuberosum, XP_012471399.1 Gossypium raimondii, XP_017618875.1 Gossypium arboretum, KAB2038530.1 Gossypium barbadense, XP_016740578.1 Gossypium hirsutum
Fig.3 Phylogenetic tree of ZAT12 homologous proteins in different species Parameters are set to be neighbor-joining,boostrap value=1 000. The numbers on the tree branches represent bootstrap confidence values as “Bootstrap” is 1000
Fig.10 Subcellular localization of GhZAT12 in tobacco leaves a,d:Confocal section of tobacco leave expressing GFP and GhZAT12-GFP,respectively. b,e:Bright field. c,f:Fluorescence and bright field merged image. Bar=50 μm
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