Biotechnology Bulletin ›› 2023, Vol. 39 ›› Issue (7): 151-159.doi: 10.13560/j.cnki.biotech.bull.1985.2022-1408
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SUN Ming-hui(), WU Qiong, LIU Dan-dan, JIAO Xiao-yu, WANG Wen-jie()
Received:
2022-11-15
Online:
2023-07-26
Published:
2023-08-17
Contact:
WANG Wen-jie
E-mail:sunminghui@aaas.org.cn;391590137@qq.com
SUN Ming-hui, WU Qiong, LIU Dan-dan, JIAO Xiao-yu, WANG Wen-jie. Cloning and Expression Analysis of CsTMFs Gene in Tea Plant[J]. Biotechnology Bulletin, 2023, 39(7): 151-159.
用途Application | 引物名称Primer name | 引物序列Primer sequence(5'-3') |
---|---|---|
实时荧光定量PCR qPCR | CsTMF1-qF | TCCATCCGTGACTCTCTTGCTGAG |
CsTMF1-qR | GCCTCCGCCTTAGTGCCTCTA | |
CsTMF2-qF | CGACTGGCATCTCTGGAATCC | |
CsTMF2-qR | GCCTCCGTCTTAATGCTTCTAACT | |
Actin-qF | GCCATCTTTGATTGGAATGG | |
Actin-qR | GGTGCCACAACCTTGATCTT | |
基因克隆Gene cloning | CsTMF1-F | ATGGCCTGGTTTAGCGGGAAAGTCTCTTTGG |
CsTMF1-R | TCATGCAGAGCCCATAGAAGAACCCATTACC | |
CsTMF2-F | ATGGCCTGGTTCGGTGGGAAAGTCT | |
CsTMF2-R | CTAAGACGAACTTAGTACCTGGATC |
Table 1 Primer sequences for PCR
用途Application | 引物名称Primer name | 引物序列Primer sequence(5'-3') |
---|---|---|
实时荧光定量PCR qPCR | CsTMF1-qF | TCCATCCGTGACTCTCTTGCTGAG |
CsTMF1-qR | GCCTCCGCCTTAGTGCCTCTA | |
CsTMF2-qF | CGACTGGCATCTCTGGAATCC | |
CsTMF2-qR | GCCTCCGTCTTAATGCTTCTAACT | |
Actin-qF | GCCATCTTTGATTGGAATGG | |
Actin-qR | GGTGCCACAACCTTGATCTT | |
基因克隆Gene cloning | CsTMF1-F | ATGGCCTGGTTTAGCGGGAAAGTCTCTTTGG |
CsTMF1-R | TCATGCAGAGCCCATAGAAGAACCCATTACC | |
CsTMF2-F | ATGGCCTGGTTCGGTGGGAAAGTCT | |
CsTMF2-R | CTAAGACGAACTTAGTACCTGGATC |
Motif | 宽度Width | 最可能序列Most possible match | 结构域Domain |
---|---|---|---|
1 | 80 | TARLQPSTVPDQSPITRRKSGFENGNLSRKLSSASSLGSMEESFFLQASLDSSDNFSERRNPGEMAMSPYFMKSMPPSAF | * |
2 | 78 | TMLVQALEELRQTLSRKEQQAVFREDMLRRDIEDLQKRYQESERRCEELVTQVPESTRPLLRQIEAMQETTGRRAEAW | * |
3 | 40 | MAWFGGKVSLGNFPDLAGAVNKISESVKNIEKNFDSALGF | * |
4 | 80 | MKMMETALQGAARQAQAKADEIAKLMNENENQKAVIEDLRRKSNEAEIESLREEYHQKVAALERKVYALTKERDTLRREQ | * |
5 | 80 | IRDSLAEELVKMTAQCEKLRVEAAILPGIRAELEALRRRHSAALELMGERDEELEELRADIVDLKEMYREQVNLLVNKIQ | TMF-TATA-bd |
6 | 79 | GLWPSGTDRKTJFEPVIAFMGHKGGESTVESIEKPELLEPPSSVEEKZEGENDRSTDFATEQTRPAEEVNZESSHMLPD | * |
7 | 79 | QAGMEVELPGFGFPTSKEIESAEEHFTDDLPDGPPPDEAAEMVSEPVSHENDTFGREVELNZQASDYEPDIKEQRVSSG | TMF-DNA-bd |
8 | 80 | VMAEGEELSKKQAAQESQIRKLRAQIRELEEEKKGLTTKLQVEENKVESIKRDKAETEKLLQETIEKHQAEJAVQKEFYT | * |
9 | 80 | QTLSRLNVLEAQISCLRAEQTQLSRSLEKERQRAAENRQEYLAAKEEADTHEGRVSQLEEEIRELRRKQKZELQEALMHR | * |
10 | 66 | TSALGPFENSELVEAKPGTNEVDQVEVGAPVPPESHNVINLHEISDEQKAZEEEIVENLSPVQAED | * |
Table 2 Detailed information of 10 motifs in the CsTMFs proteins
Motif | 宽度Width | 最可能序列Most possible match | 结构域Domain |
---|---|---|---|
1 | 80 | TARLQPSTVPDQSPITRRKSGFENGNLSRKLSSASSLGSMEESFFLQASLDSSDNFSERRNPGEMAMSPYFMKSMPPSAF | * |
2 | 78 | TMLVQALEELRQTLSRKEQQAVFREDMLRRDIEDLQKRYQESERRCEELVTQVPESTRPLLRQIEAMQETTGRRAEAW | * |
3 | 40 | MAWFGGKVSLGNFPDLAGAVNKISESVKNIEKNFDSALGF | * |
4 | 80 | MKMMETALQGAARQAQAKADEIAKLMNENENQKAVIEDLRRKSNEAEIESLREEYHQKVAALERKVYALTKERDTLRREQ | * |
5 | 80 | IRDSLAEELVKMTAQCEKLRVEAAILPGIRAELEALRRRHSAALELMGERDEELEELRADIVDLKEMYREQVNLLVNKIQ | TMF-TATA-bd |
6 | 79 | GLWPSGTDRKTJFEPVIAFMGHKGGESTVESIEKPELLEPPSSVEEKZEGENDRSTDFATEQTRPAEEVNZESSHMLPD | * |
7 | 79 | QAGMEVELPGFGFPTSKEIESAEEHFTDDLPDGPPPDEAAEMVSEPVSHENDTFGREVELNZQASDYEPDIKEQRVSSG | TMF-DNA-bd |
8 | 80 | VMAEGEELSKKQAAQESQIRKLRAQIRELEEEKKGLTTKLQVEENKVESIKRDKAETEKLLQETIEKHQAEJAVQKEFYT | * |
9 | 80 | QTLSRLNVLEAQISCLRAEQTQLSRSLEKERQRAAENRQEYLAAKEEADTHEGRVSQLEEEIRELRRKQKZELQEALMHR | * |
10 | 66 | TSALGPFENSELVEAKPGTNEVDQVEVGAPVPPESHNVINLHEISDEQKAZEEEIVENLSPVQAED | * |
物种名Species name | 登录号Accession No. |
---|---|
水稻Oryza sativa Japonica Group | BAG90311.1 |
茭白Zizania palustris | KAG8086397 |
小麦Triticum aestivum | XP 044346450.1 |
大麦Hordeum vulgare subsp. Vulgar | BAJ99499 |
玉米Zea mays | XP 008656031.1 |
Table 3 Accession numbers of different species of TMF
物种名Species name | 登录号Accession No. |
---|---|
水稻Oryza sativa Japonica Group | BAG90311.1 |
茭白Zizania palustris | KAG8086397 |
小麦Triticum aestivum | XP 044346450.1 |
大麦Hordeum vulgare subsp. Vulgar | BAJ99499 |
玉米Zea mays | XP 008656031.1 |
Fig. 8 Multiple sequence alignment of tea plant TMF with other plants The conserved domain of TMF-DNA-bd is in purple box, and the conserved domain of TMF-TATA-bd is in black box
Fig. 10 Expression pattern of CsTMFs gene under drought stress and low temperature Different lowercase letters in figure indicate significant differences at different time of the same treatment(P<0.05)
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