At5g66750(DDM1) ORF Clone

产品内容

pLinker1-At5g66750------------------------2 μg

TE buffer--------------------------------------500 μl

产品信息

产品名称：At5g66750(DDM1) ORF Clone

产品货号：QA5g66750

别称：DDM1;CHA1;CHR1;SOM1;SOM4;At5g66750;MSN2.14

产品类型：ORF克隆载体

基因ID：At5g66750

转录本：AT5G66750.1

载体：pLinker1

抗性：kanR

ORF 长度：2295 bp

蛋白长度：764 aa

蛋白分子量：86638 Da

片段物种：拟南芥（Arabidopsis thaliana）

NCBI gene ID：836808

Uniprot ID：Q9XFH4

基因摘要

Protein is similar to SWI2/SNF2 chromatin remodeling proteins. DDM1 is appears to act as a chromatin-remodeling ATPase involved in cytosine methylation in CG and non-CG contexts. Involved in gene silencing and maintenance of DNA methylation and histone methylation. Hypomethylation of many genomic regions occurs in ddm1 mutants, and can cause several phenotypic abnormalities, but some loci, such as BONSAI (At1g73177) can be hypermethylated in ddm1 mutants after several generations, leading to different phenotypes. DDM1 might be involved in establishing a heterochromain boundary. A line expressing an RNAi targeted against DDM1 shows some resistance to agrobacterium-mediated root transformation.

ORF 核苷酸序列

>At5g66750(DDM1) ORF Clone

ACGCTAGCATGGATGTTTTCCCAGTCACGACGTTGTAAAACGACGGCCAGTCTTAAGCTCCAGTGAATTCGAGCTCGGTACCTCGCGAATGCATCTAGATATC

ATGGTTAGTCTGCGCTCCAGAAAAGTTATTCCGGCTTCGGAAATGGTCAGCGACGGGAAAACGGAGAAAGATGCGTCTGGTGATTCACCCACTTCTGTTCTCAACGAAGAGGAAAACTGTGAGGAGAAAAGTGTTACTGTTGTAGAGGAAGAGATACTTCTAGCCAAAAATGGAGATTCTTCTCTTATTTCTGAAGCCATGGCTCAGGAGGAAGAGCAGCTGCTCAAACTTCGGGAAGATGAAGAGAAAGCTAACAATGCTGGATCTGCTGTTGCTCCTAATCTGAATGAAACTCAGTTTACTAAACTTGATGAGCTCTTGACGCAAACTCAGCTCTACTCTGAGTTTCTCCTTGAGAAAATGGAGGATATCACAATTAATGGGATAGAAAGTGAGAGCCAAAAAGCTGAGCCCGAGAAGACTGGTCGTGGACGCAAAAGAAAGGCTGCTTCTCAGTACAACAATACTAAGGCTAAGAGAGCGGTTGCTGCTATGATTTCAAGATCTAAAGAAGATGGTGAGACCATCAACTCAGATCTGACAGAGGAAGAAACAGTCATCAAACTGCAGAATGAACTTTGTCCTCTTCTCACTGGTGGACAGTTAAAGTCTTATCAGCTTAAAGGTGTCAAATGGCTAATATCATTGTGGCAGAATGGTTTGAATGGAATATTAGCTGATCAAATGGGACTTGGAAAGACGATTCAAACGATCGGTTTCTTATCACATCTGAAAGGGAATGGGTTGGATGGTCCATATCTAGTCATTGCTCCACTGTCTACACTTTCAAATTGGTTCAATGAGATTGCTAGGTTCACGCCTTCCATCAATGCAATCATCTACCATGGGGATAAAAATCAAAGGGATGAGCTCAGGAGGAAGCACATGCCTAAAACTGTTGGTCCCAAGTTCCCTATAGTTATTACTTCTTATGAGGTTGCCATGAATGATGCTAAAAGAATTCTGCGGCACTATCCATGGAAATATGTTGTGATTGATGAGGGCCACAGGTTGAAAAACCACAAGTGTAAATTGTTGAGGGAACTAAAACACTTGAAGATGGATAACAAACTTCTGCTGACAGGAACACCTCTGCAAAATAATCTTTCTGAGCTTTGGTCTTTGTTAAATTTTATTCTGCCTGACATCTTTACATCACATGATGAATTTGAATCATGGTTTGATTTTTCTGAAAAGAACAAAAACGAAGCAACCAAGGAAGAAGAAGAGAAAAGAAGAGCTCAAGTTGTTTCCAAACTTCATGGTATACTACGACCATTCATCCTTCGAAGAATGAAATGTGATGTTGAGCTCTCACTTCCACGGAAAAAGGAGATTATAATGTATGCTACAATGACTGATCATCAGAAAAAGTTCCAGGAACATCTGGTGAATAACACGTTGGAAGCACATCTTGGAGAGAATGCCATCCGAGGTCAAGGCTGGAAGGGAAAGCTTAACAACCTGGTCATTCAACTTCGAAAGAACTGCAACCATCCTGACCTTCTCCAGGGGCAAATAGATGGTTCATATCTCTACCCTCCTGTTGAAGAGATTGTTGGACAGTGTGGTAAATTCCGCTTATTGGAGAGATTACTTGTTCGGTTATTTGCCAATAATCACAAAGTCCTTATCTTCTCCCAATGGACGAAACTTTTGGACATTATGGATTACTACTTCAGTGAGAAGGGGTTTGAGGTTTGCAGAATCGATGGCAGTGTGAAGCTGGATGAAAGGAGAAGACAGATTAAAGATTTCAGTGATGAGAAGAGCAGCTGTAGTATATTTCTCCTGAGTACCAGAGCTGGAGGACTCGGAATCAATCTTACTGCTGCTGATACATGCATCCTCTATGACAGCGACTGGAACCCTCAAATGGACTTGCAAGCCATGGACAGATGCCACAGAATCGGGCAGACGAAACCTGTTCATGTTTATAGGCTTTCCACGGCTCAGTCGATAGAGACCCGGGTTCTGAAACGAGCGTACAGTAAGCTCAAGCTGGAACATGTGGTTATTGGCCAAGGGCAGTTTCATCAAGAACGTGCCAAGTCTTCAACACCTTTAGAGGAAGAGGACATACTGGCGTTGCTTAAGGAAGATGAAACTGCTGAAGATAAGTTGATACAAACCGATATAAGCGATGCGGATCTTGACAGGTTACTTGACCGGAGTGACCTGACAATTACTGCACCGGGAGAGACACAAGCTGCTGAAGCTTTTCCAGTGAAGGGTCCAGGTTGGGAAGTGGTCCTGCCTAGTTCGGGAGGAATGCTGTCTTCCCTGAACAGTTAG

GATATCGGATCCCGGGCCCGTCGACTGCAGAGGCCTGCATGCAAGCTTGGCGTAATCATGGTCATACCCTATAGTGAGTCGTATTACATGGTCATAGCTG

黑色：载体序列、红色：同源重组序列、绿色：EcoR V酶切位点、蓝色：ORF序列

蛋白简介

ATP-dependent DNA helicase that plays a role in formation, organization, stability and heritability of heterochromatin and thus regulates several physiological traits. Binds to the nucleosome and promotes chromatin remodeling in an ATP-dependent manner; induces nucleosome repositioning on a short DNA fragment, and, possibly, could be guided to target sites (including silent transposable elements) by small interfering RNAs (siRNAs). Can bind both free and nucleosomal DNA. Required for the heritable maintenance of genome integrity and transcriptional gene silencing (TGS), including homology-dependent gene silencing (HDG silencing), via the maintenance of DNA methylation (mostly on cytosine, in both CpG and CpHpG sites, where H is A, T or C) and of histone methylation (e.g. chromatin methylation). May facilitate localization of MBD proteins at specific nuclear domains. Necessary for the maintenance of the genomic imprint at the MEA locus, especially for the silencing of paternally inherited MEA locus. Plays a major role in the inactivation maintenance of retrotransposons (e.g. Tar17, SINE, LINE, ATLN39, CAC1 (CACTAs), Athila elements, and mutator-like elements MULEs and TIR-MULEs) and the silencing of repeated genes and transgenes (e.g. T-DNA insertions). Required for KYP-dependent histone H3 'Lys-9' (H3K9me) methylation, deacetylation of histone H4 'Lys-16' (H4K16) and MET1-dependent DNA methylation. Involved in the chromatin organization of 5S rRNA genes (localized in the pericentromeric heterochromatin of chromosomes 3, 4, and 5) modifications during heterochromatin establishment. Prevents siRNA accumulation (siRNA are probably involved in epigenetic inheritance and in 5S rRNA genes regulation by silencing). Required during plant organogenesis and development, as well as during seed formation. {ECO:0000269|PubMed:10319870, ECO:0000269|PubMed:10580004, ECO:0000269|PubMed:10715322, ECO:0000269|PubMed:10939915, ECO:0000269|PubMed:11148284, ECO:0000269|PubMed:11238379, ECO:0000269|PubMed:11346800, ECO:0000269|PubMed:11779837, ECO:0000269|PubMed:11948614, ECO:0000269|PubMed:12077425, ECO:0000269|PubMed:12194816, ECO:0000269|PubMed:12370435, ECO:0000269|PubMed:12403775, ECO:0000269|PubMed:12456661, ECO:0000269|PubMed:12472695, ECO:0000269|PubMed:12609046, ECO:0000269|PubMed:12663548, ECO:0000269|PubMed:14608503, ECO:0000269|PubMed:14630972, ECO:0000269|PubMed:14691539, ECO:0000269|PubMed:15269773, ECO:0000269|PubMed:15514067, ECO:0000269|PubMed:15516340, ECO:0000269|PubMed:15805479, ECO:0000269|PubMed:15968513, ECO:0000269|PubMed:17012404, ECO:0000269|PubMed:17082818, ECO:0000269|PubMed:17144899, ECO:0000269|PubMed:17339215, ECO:0000269|PubMed:17627280, ECO:0000269|PubMed:18467467, ECO:0000269|PubMed:19529764, ECO:0000269|PubMed:7870578, ECO:0000269|PubMed:8316832, ECO:0000269|PubMed:8901594, ECO:0000269|PubMed:9450349, ECO:0000269|PubMed:9611181, ECO:0000269|PubMed:9620857, ECO:0000269|PubMed:9927473}.

蛋白质序列

>ATP-dependent DNA helicase DDM1 (EC 3.6.4.12) (Protein CHROMATIN REMODELING 1) (AtCHR1) (CHR01) (Protein DECREASED DNA METHYLATION 1) (AtDDM1) (Protein SOMNIFEROUS 1) (SWI/SNF2-related matrix-associated actin-dependent regulator of chromatin DDM1)

MVSLRSRKVIPASEMVSDGKTEKDASGDSPTSVLNEEENCEEKSVTVVEEEILLAKNGDSSLISEAMAQEEEQLLKLREDEEKANNAGSAVAPNLNETQFTKLDELLTQTQLYSEFLLEKMEDITINGIESESQKAEPEKTGRGRKRKAASQYNNTKAKRAVAAMISRSKEDGETINSDLTEEETVIKLQNELCPLLTGGQLKSYQLKGVKWLISLWQNGLNGILADQMGLGKTIQTIGFLSHLKGNGLDGPYLVIAPLSTLSNWFNEIARFTPSINAIIYHGDKNQRDELRRKHMPKTVGPKFPIVITSYEVAMNDAKRILRHYPWKYVVIDEGHRLKNHKCKLLRELKHLKMDNKLLLTGTPLQNNLSELWSLLNFILPDIFTSHDEFESWFDFSEKNKNEATKEEEEKRRAQVVSKLHGILRPFILRRMKCDVELSLPRKKEIIMYATMTDHQKKFQEHLVNNTLEAHLGENAIRGQGWKGKLNNLVIQLRKNCNHPDLLQGQIDGSYLYPPVEEIVGQCGKFRLLERLLVRLFANNHKVLIFSQWTKLLDIMDYYFSEKGFEVCRIDGSVKLDERRRQIKDFSDEKSSCSIFLLSTRAGGLGINLTAADTCILYDSDWNPQMDLQAMDRCHRIGQTKPVHVYRLSTAQSIETRVLKRAYSKLKLEHVVIGQGQFHQERAKSSTPLEEEDILALLKEDETAEDKLIQTDISDADLDRLLDRSDLTITAPGETQAAEAFPVKGPGWEVVLPSSGGMLSSLNS

关键词

ATP-binding;Coiled coil;DNA-binding;Helicase;Hydrolase;Nucleotide-binding;Nucleus;Reference proteome;Repeat;Transcription;Transcription regulation


质粒图谱

产品说明

本产品是经过离子交换柱纯化后，装在无菌冻存管中通过真空干燥获得的2 μg连有At5g66750(DDM1) 基因ORF的质粒DNA干粉。

保存：-20 ℃

使用说明

1、以5000 × g离心5 min；

2、打开冻存管，加入20 μL无菌水溶解DNA，并在室温下孵育10 min；

3、短暂涡旋冻存管，然后瞬时离心 (小于5000 × g)。

4、质粒溶液在-20℃下可稳定保存一年，请避免反复冻融。


扩增方案

1、收到质粒干粉后请先5000 rpm离心1 min，再加入20 μL无菌水溶解质粒，室温放置1 min；

2、从-80 °C冰箱中取出相应的感受态，置于冰盒上解冻，并做好标记；

3、取2 μL质粒加至100 μL感受态中，冰浴30min;

4、42 °C热激90 s，再冰浴2 min;

5、加入900 μL无抗的LB液体培养基，200 rpm震荡培养45 min;

6、5000 rpm离心5 min，仅留100 μL上清混匀菌体沉淀；

7、混匀后的菌液加至对应抗性的LB平板上，倒入适量玻璃珠，涂匀液体；

8、将平板正向培养1 h，再倒置培养12 h～16 h;

9、挑取单克隆菌落至对应抗性的LB液体培养基中，震荡培养12 h~16 h，根据实验需要提取质粒。

注意事项

1、同一个基因的单个转录序列可能由于发生自然变异 (例如核苷酸多态性)而有所不同，每一种都有其自身有效的存在。

2、该克隆被工程化，但表达取决于基因的性质。

3、本产品仅限于科学研究，不得用于临床诊断或治疗，不得用于食品或药品。

4、为了您的安全和健康，请穿实验服并戴一次性手套操作。

Tair 网站：https://www.arabidopsis.org/servlets/Search?type=general&search_action=detail&method=1&show_obsolete=F&name=At5g66750&sub_type=gene&SEARCH_EXACT=4&SEARCH_CONTAINS=1
NCBI网站：https://www.ncbi.nlm.nih.gov/gene/?term=836808
Uniprot 网站：https://www.uniprot.org/uniprotkb//entry

相关文献 (NCBI PubMed ID)

PMID: 10319870; 10718197; 27862469; 14593172; 15516340; 8316832; 7870578; 8901594; 9450349; 9611181; 9620857; 10939915; 9927473; 10580004; 10715322; 11148284; 10999404; 11238379; 11346800; 11948614; 12194816; 12456661; 11779837; 12472695; 12370435; 12077425; 12663548; 12403775; 14630972; 12609046; 14691539; 12597869; 15514067; 14608503; 15269773; 15968513; 15805479; 17082818; 16547115; 17012404; 17627280; 17339215; 17144899; 18467467; 19529764; 24265739