应用SELEX技术检测水稻RFL转录因子的DNA结合位点

doi:10.11983/CBB16130

植物学报 ›› 2017, Vol. 52 ›› Issue (1): 70-76.DOI: 10.11983/CBB16130 cstr: 32102.14.CBB16130

所属专题：水稻生物学专辑 (2017年52卷1期)

应用SELEX技术检测水稻RFL转录因子的DNA结合位点

丁承强, 常忠原, 王琰, 王绍华, 丁艳锋^*()

南京农业大学农学院, 南京 210095

收稿日期:2016-06-13 接受日期:2016-10-13 出版日期:2017-01-01 发布日期:2017-01-23
通讯作者: 丁艳锋
作者简介:
# 共同第一作者
基金资助:
国家自然科学基金(No.31401324)

Use of SELEX for Research of the DNA Binding Sites of the RFL, a Transcription Factor of Rice

Chengqiang Ding, Zhongyuan Chang, Yan Wang, Shaohua Wang, Yanfeng Ding^*

College of Agronomy, Nanjing Agricultural University, Nanjing 210095, China

Received:2016-06-13 Accepted:2016-10-13 Online:2017-01-01 Published:2017-01-23
Contact: Ding Yanfeng
About author:
# Co-first authors

摘要/Abstract

摘要： FLORICAULA/LEAFY是植物特有的转录因子, 其可以通过调控下游基因的表达对植物的发育起到调控作用。水稻(Oryza sativa)中的同源基因RFL (Rice FLORICAULA/LEAFY)能够调控多个发育过程, 包括开花、叶间期以及枝梗和颖花分化。指数富集配体的系统进化(SELEX)技术已经被广泛应用于体外筛选转录因子的DNA结合位点。应用SELEX技术检测水稻RFL的DNA结合序列, 经过7次PCR循环富集过程获得结合特异序列, 通过测序和MEME分析, 鉴定到RFL的DNA结合序列为(C/A)(C/T)NN(T/C/A)G(G/T)。实验结果为进一步阐明RFL分子功能奠定了基础。

Abstract: FLORICAULA/LEAFY encodes an evolutionarily conserved land plant-specific transcription factor. It functions by regulating the expression of downstream genes. RFL, the FLORICAULA/LEAFY homolog in rice, controls several important developments in rice: flowering time, plastochron and panicle branching. In this study, we detected the binding specificity of RFL by using the method systematic evolution of ligands by exponential enrichment (SELEX). After seven SELEX processes and sequencing, the binding site of RFL was identified as (C/A)(C/T)NN(T/C/A)G(G/T). The result contributes to clarifying the molecular function of RFL.

丁承强, 常忠原, 王琰, 王绍华, 丁艳锋. 应用SELEX技术检测水稻RFL转录因子的DNA结合位点. 植物学报, 2017, 52(1): 70-76.

Chengqiang Ding, Zhongyuan Chang, Yan Wang, Shaohua Wang, Yanfeng Ding. Use of SELEX for Research of the DNA Binding Sites of the RFL, a Transcription Factor of Rice. Chinese Bulletin of Botany, 2017, 52(1): 70-76.

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图/表 4

参考文献 17

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Number	Sequences (5'-3')	Number	Sequences (5'-3')
1	TGTGACCGCGGGATAAACGAGCTGTC	42	CGACCCGGGGATCGCGTTTGTACTC
2	CCCGGTGGGCAGAATCCGATCGGACC	43	CCCCCCAAGGGTAATAACATGGGCGG
3	TGTAACCGGGTTTACCCATAGGTTAC	44	CCGGCACCCCGGACATACCCAATGGT
4	CAAACCCTGTGTGTAATAACCGC	45	GACCGGGTATTCCCGACAAACCAGTA
5	GGCCACGGGGTTTACTCGGCAGCATG	46	AGACCGGTGGCCGGCGCCTAGAAACC
6	CGTTATACTAGATGTCCAGCGGGTAC	47	GTTCAGGCGGCCGTGTAATGTCCGCT
7	CGGACTCGGGTTAATACGGTGGGTCC	48	CCCTGAGACGAGGTGTTTTGAGGTCC
8	GGTGTCGAGTGGTATTCCACCGGTCC	49	CGGACCCATGTAATACCCGCGTAATG
9	GCCCTTTGGGTTTTGTCCCCCGTATA	50	CAACCACCAATGGTCCATACGGGCGG
10	TGAGACTACAGCGGGTAATAGCCTTA	51	TCCAAAGGTCAGTCACCGGGTCTGCG
11	GGTCCTGCCTGCTAACCCCGTGGTCC	52	GTGCATGGACCGACTGTCAACGGAAA
12	GGGTCCACAGGCAGGTCGTTGTACAT	53	GAGACTTTTGCCTCAAGTGTGGTCC
13	AGACGGTCCGGTAGCTGCTTTCTTGC	54	AAAGTCCAAAGCGGGGGGGCAGACCG
14	GTGTGGGGCGGGTTCTGGGCGGACAA	55	CCGAAACCCCTCCGGTCACAATCCC
15	CGCCGTACACTAAGGACCGCTGGCTA	56	CGGTACCGGGTATTGACCGAAGGTGC
16	CGGTAATTGTTGAACGATCCGGGCTG	57	TCCGTAGGTGGTATCCCCAGTGTGTA
17	GCGACCGGTTGCTCGGTTGTGGTCCC	58	GACGCCAGGGGTTATTACCTGTGCCG
18	GTTTAGGGTTTAGTACCCGTCGGTCC	59	TCCTATATCCTAATGGTGGAGGACCG
19	TGGCTAATGTCCCCGTGGCTCCCCGC	60	TTACCCAAACGGTAATAGCACGGACC
20	CATGATAGGTTTTAGCTGAGGGGCCA	61	CAACTATACCCAACGAGCATAACCCG
21	ACCGTTGGTTCCTCGCTTTTCTTAGC	62	TGACTACTCGGCAAGTCTTTACCCTG
22	TTCGAACGCGTGCAAGGACCGGTGGA	63	CCAGAAGTACTTATCCAGGGTAATAA
23	GGAGCTGGACTTTACCCTTGCTTTGC	64	CCTGGGCTTTGCATTTCGGTCCTGGA
24	GAGTTAAATGATTATCCCGTGGTCCC	65	GACCTGGACTATAACACTGGACCGTA
25	TGACCGATGGGGTGCAAGCTGAGGGA	66	GCGTGGACCGGTCCGTTACTCCGTCC
26	GGGTGCTACCACCGGCTGGGTTCGGG	67	CGAGGTAGCGACACGACTGGTGGTCC
27	CGGGATGAGCGAGTACGAGTGGGCCC	68	CCGAAAGGACCGTAACCCAACGGTTA
28	TCCCTCAGGGTATTACCCCTATGGCC	69	CGACTACAACGACCCCACGGGTAAAG
29	TGGGCATTACCTTGAACCACCTGGTT	70	GAGGCCGCGAGGCCCAGACCAATGGA
30	CACCAATGGGTATTGTCTGTAGCCC	71	TGTGCTTTTGTCGCTTCGCCCCACGG
31	CCCGCCGGGTAATTGTTGTCGTAACG	72	ACCGATGGACCGCCGTGGGGTTGTCG
32	GTCTTTTGGTCCGTGTCTTGGCGGGG	73	ACCTCCTGGCTATTGTACTCCGGTTA
33	CGGGTAAAGGGGCTTGGTCCCCCAGC	74	CCCGTCGGTATTTAGCTGGTAGTCG
34	TGTGAGGACGTGGTTGGACCGGTCTG	75	ACTGGTTCACTGCCGGTATACGGTCA
35	ACCGACCGTTGGGCTTAATCCTCTTG	76	GAAACCTGCATTTGTCCACTGGTCTA
36	CTCCAAATGCACCCAAGCGGGTTCCA	77	CACCGAGCGATAAAACCGTTGGTCTA
37	CCCAAAACCCACCGGTCGTTGCCCCG	78	AGTGGTGGTTATTGCCCAATGTGCCC
38	AGGGCTTAACCTCTTTATGGACCCCG	79	GGACATTAACCGGCCAACTCCGCGGA
39	GGACCACAGGGTAAGGCTCATCGCTA	80	GATCCGCTGGGTAACCCAGGAAGGGT
40	GCGACCAGGCAAAGACTCCTGTACGC	81	ACCGTCGGTTGCAGGTCCTTGGTCCG
41	AGTCATACGCGGCTATTACCTTCCGG

Number	Sequences (5'-3')	Number	Sequences (5'-3')
1	TGTGACCGCGGGATAAACGAGCTGTC	42	CGACCCGGGGATCGCGTTTGTACTC
2	CCCGGTGGGCAGAATCCGATCGGACC	43	CCCCCCAAGGGTAATAACATGGGCGG
3	TGTAACCGGGTTTACCCATAGGTTAC	44	CCGGCACCCCGGACATACCCAATGGT
4	CAAACCCTGTGTGTAATAACCGC	45	GACCGGGTATTCCCGACAAACCAGTA
5	GGCCACGGGGTTTACTCGGCAGCATG	46	AGACCGGTGGCCGGCGCCTAGAAACC
6	CGTTATACTAGATGTCCAGCGGGTAC	47	GTTCAGGCGGCCGTGTAATGTCCGCT
7	CGGACTCGGGTTAATACGGTGGGTCC	48	CCCTGAGACGAGGTGTTTTGAGGTCC
8	GGTGTCGAGTGGTATTCCACCGGTCC	49	CGGACCCATGTAATACCCGCGTAATG
9	GCCCTTTGGGTTTTGTCCCCCGTATA	50	CAACCACCAATGGTCCATACGGGCGG
10	TGAGACTACAGCGGGTAATAGCCTTA	51	TCCAAAGGTCAGTCACCGGGTCTGCG
11	GGTCCTGCCTGCTAACCCCGTGGTCC	52	GTGCATGGACCGACTGTCAACGGAAA
12	GGGTCCACAGGCAGGTCGTTGTACAT	53	GAGACTTTTGCCTCAAGTGTGGTCC
13	AGACGGTCCGGTAGCTGCTTTCTTGC	54	AAAGTCCAAAGCGGGGGGGCAGACCG
14	GTGTGGGGCGGGTTCTGGGCGGACAA	55	CCGAAACCCCTCCGGTCACAATCCC
15	CGCCGTACACTAAGGACCGCTGGCTA	56	CGGTACCGGGTATTGACCGAAGGTGC
16	CGGTAATTGTTGAACGATCCGGGCTG	57	TCCGTAGGTGGTATCCCCAGTGTGTA
17	GCGACCGGTTGCTCGGTTGTGGTCCC	58	GACGCCAGGGGTTATTACCTGTGCCG
18	GTTTAGGGTTTAGTACCCGTCGGTCC	59	TCCTATATCCTAATGGTGGAGGACCG
19	TGGCTAATGTCCCCGTGGCTCCCCGC	60	TTACCCAAACGGTAATAGCACGGACC
20	CATGATAGGTTTTAGCTGAGGGGCCA	61	CAACTATACCCAACGAGCATAACCCG
21	ACCGTTGGTTCCTCGCTTTTCTTAGC	62	TGACTACTCGGCAAGTCTTTACCCTG
22	TTCGAACGCGTGCAAGGACCGGTGGA	63	CCAGAAGTACTTATCCAGGGTAATAA
23	GGAGCTGGACTTTACCCTTGCTTTGC	64	CCTGGGCTTTGCATTTCGGTCCTGGA
24	GAGTTAAATGATTATCCCGTGGTCCC	65	GACCTGGACTATAACACTGGACCGTA
25	TGACCGATGGGGTGCAAGCTGAGGGA	66	GCGTGGACCGGTCCGTTACTCCGTCC
26	GGGTGCTACCACCGGCTGGGTTCGGG	67	CGAGGTAGCGACACGACTGGTGGTCC
27	CGGGATGAGCGAGTACGAGTGGGCCC	68	CCGAAAGGACCGTAACCCAACGGTTA
28	TCCCTCAGGGTATTACCCCTATGGCC	69	CGACTACAACGACCCCACGGGTAAAG
29	TGGGCATTACCTTGAACCACCTGGTT	70	GAGGCCGCGAGGCCCAGACCAATGGA
30	CACCAATGGGTATTGTCTGTAGCCC	71	TGTGCTTTTGTCGCTTCGCCCCACGG
31	CCCGCCGGGTAATTGTTGTCGTAACG	72	ACCGATGGACCGCCGTGGGGTTGTCG
32	GTCTTTTGGTCCGTGTCTTGGCGGGG	73	ACCTCCTGGCTATTGTACTCCGGTTA
33	CGGGTAAAGGGGCTTGGTCCCCCAGC	74	CCCGTCGGTATTTAGCTGGTAGTCG
34	TGTGAGGACGTGGTTGGACCGGTCTG	75	ACTGGTTCACTGCCGGTATACGGTCA
35	ACCGACCGTTGGGCTTAATCCTCTTG	76	GAAACCTGCATTTGTCCACTGGTCTA
36	CTCCAAATGCACCCAAGCGGGTTCCA	77	CACCGAGCGATAAAACCGTTGGTCTA
37	CCCAAAACCCACCGGTCGTTGCCCCG	78	AGTGGTGGTTATTGCCCAATGTGCCC
38	AGGGCTTAACCTCTTTATGGACCCCG	79	GGACATTAACCGGCCAACTCCGCGGA
39	GGACCACAGGGTAAGGCTCATCGCTA	80	GATCCGCTGGGTAACCCAGGAAGGGT
40	GCGACCAGGCAAAGACTCCTGTACGC	81	ACCGTCGGTTGCAGGTCCTTGGTCCG
41	AGTCATACGCGGCTATTACCTTCCGG

应用SELEX技术检测水稻RFL转录因子的DNA结合位点

Use of SELEX for Research of the DNA Binding Sites of the RFL, a Transcription Factor of Rice

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