At5g60300(LECRK19) ORF Clone

产品内容

pLinker1-At5g60300------------------------2 μg

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

产品信息

产品名称：At5g60300(LECRK19) ORF Clone

产品货号：QA5g60300

别称：LECRK19;DORN1;P2K1;At5g60300;F15L12.17

产品类型：ORF克隆载体

基因ID：At5g60300

转录本：AT5G60300.3

载体：pLinker1

抗性：kanR

ORF 长度：2301 bp

蛋白长度：766 aa

蛋白分子量：85319 Da

片段物种：拟南芥（Arabidopsis thaliana）

NCBI gene ID：836152

Uniprot ID：Q9LSR8

基因摘要

Encodes a legume-type lectin receptor kinase that is structurally distinct from the mammalian extracellular ATP receptors and acts as an extracellular ATP receptor in Arabidopsis. Extracellular ATP acts as a damage-associated molecular pattern in plants, and its signaling through P2K1 is important for mounting an effective defense response against various pathogenic microorganisms. It also plays a role in cell wall-plasma membrane adhesion.

ORF 核苷酸序列

>At5g60300(LECRK19) ORF Clone

ACGCTAGCATGGATGTTTTCCCAGTCACGACGTTGTAAAACGACGGCCAGTCTTAAGCTCCAGTGAATTCGAGCTCGGTACCTCGCGAATGCATCTAGATATC

ATGGCTCGTTGGTTGCTTCAGATCCTGATCATCTCTTCTCTTCATCTGAGCTCTGTATCAAGTCAACAAGAGACAAGCTTTGTCTATGAAAGCTTCCTCGACCGACAAAATCTTTATCTTGACAAATCTGCAATAGTACTTCCCAGTGGATTATTGCAGTTGACAAATGCTTCAGAGCACCAGATGGGTCATGCTTTCCACAAGAAACCAATTGAGTTCAGTTCATCTGGACCACTCTCCTTCTCAACACACTTTGTGTGTGCTTTGGTGCCTAAGCCAGGTTTTGAAGGTGGCCATGGTATTGTCTTTGTATTGTCTCCTTCTATGGACTTTACTCACGCAGAATCAACTAGATACTTGGGGATTTTCAATGCTTCAACAAATGGATCTTCCTCTTATCACGTACTTGCTGTTGAGCTTGACACTATTTGGAACCCAGATTTCAAAGATATTGACCACAATCATGTGGGGATTGATGTGAACAGTCCTATATCTGTCGCAATAGCTTCAGCATCTTACTATTCCGACATGAAAGGGAGTAATGAAAGCATAAACCTCTTGAGTGGAAACCCTATACAGGTCTGGGTGGATTATGAAGGCACTTTGCTCAATGTTTCGGTTGCTCCTCTTGAAGTCCAGAAGCCAACTCGACCTCTTTTATCACACCCCATCAACCTTACAGAACTTTTTCCAAATAGATCAAGTCTGTTTGCTGGGTTCTCTGCAGCAACAGGGACAGCGATCAGTGATCAATATATTCTCTGGTGGAGTTTTAGCATAGACAGAGGATCGCTGCAGCGACTTGATATCTCAAAACTTCCTGAAGTTCCTCATCCTAGAGCTCCACATAAGAAAGTATCTACACTGATTATTCTCCTACCCGTTTGTCTGGCTATTTTGGTGTTGGCTGTTCTTGCAGGACTTTATTTCCGCAGGAGACGCAAGTATTCAGAAGTATCTGAAACATGGGAAAAGGAGTTTGATGCGCATCGGTTCTCTTACAGGTCCTTGTTCAAAGCAACAAAGGGGTTCAGCAAAGATGAATTTCTGGGAAAAGGAGGTTTTGGAGAAGTTTATAGAGGAAATCTCCCACAAGGCAGAGAAATAGCAGTGAAAAGAGTGTCCCATAATGGTGATGAAGGTGTGAAGCAGTTTGTGGCTGAAGTCGTGAGCATGAGATGTCTGAAACACAGGAATCTTGTACCACTGTTTGGGTATTGCAGAAGAAAACGAGAACTTCTTCTTGTCTCAGAGTACATGCCCAATGGTAGTCTTGACGAGCATTTGTTTGATGACCAGAAACCGGTTCTTTCTTGGTCACAGAGACTTGTGGTCGTTAAGGGTATAGCCTCTGCTCTCTGGTACCTGCATACAGGTGCTGACCAAGTTGTTCTGCACCGAGATGTCAAAGCTTCCAACATTATGTTAGACGCGGAGTTCCATGGAAGGTTAGGGGATTTTGGCATGGCCAGGTTTCATGAGCACGGAGGCAATGCAGCTACAACAGCAGCTGTTGGGACTGTAGGGTACATGGCCCCGGAGCTAATCACAATGGGAGCTTCCACTGGAACTGATGTTTACGCATTTGGTGTTTTCATGCTTGAAGTAACCTGTGGGAGGAGACCCGTGGAACCGCAGTTGCAAGTTGAGAAACGACATATGATCAAATGGGTTTGTGAGTGCTGGAAAAAGGATTCTTTGCTTGATGCTACTGACCCGAGATTGGGAGGTAAATTTGTAGCTGAGGAAGTCGAGATGGTTATGAAACTGGGTCTGCTCTGTTCAAATATAGTGCCAGAATCCAGACCCACAATGGAACAAGTGGTCCTGTACCTAAACAAAAACCTACCTTTGCCAGATTTCTCACCATACACTTTGGGGATTGGCACATTTGCTCCGGTTCTGGTAGATGCATCCTCCCTTGTAGTCTCTTCCGCTTCATGGAGTTTGTCAGGTCCATCGATGTCTTCATCCTCGCCCAACCACTCACCTTACGCTTGGCAGTCAACAGATCAGCCATGGGGACAGACAATAGACACTAAGAACTCTCTTCACATAGTTGCAGAACCAGAGAAGCCCTCACCCGCTGTCAAAATGGTCACATTGCCTGCAGAAGATCCTCAGTCCAATCATTCCAGCATCAGCAGTCAGAGAGTCCAGCCTGTAAAGAGGGAAAAACGGCGACTGCATCAGATACTGGTGGCATTCCCTTGGATAAACAAACAATACTTTAAACTTGGATTACCAAAACATATTGTACATGTTTCTTTGTTCTTTTTTCTTCAGCTTGCGAGGTTATGA

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

Essential receptor for extracellular ATP (PubMed:24436418, PubMed:25301072). Binds ATP with high affinity through its extracellular legume-lectin like region (PubMed:24436418). Is required for ATP-induced intracellular calcium response, mitogen-activated protein kinase 3 (MPK3) and MPK6 activation and ATP-induced gene expression (PubMed:24436418, PubMed:25301072). May play a variety of roles in stress resistance (PubMed:24436418). May be involved in protein-protein interactions with RGD motif-containing proteins as potential ligands (PubMed:21483488, PubMed:16361528). Plays probably a structural and signaling role at the plant cell surfaces (PubMed:24436418, PubMed:21483488). {ECO:0000269|PubMed:16361528, ECO:0000269|PubMed:21483488, ECO:0000269|PubMed:24436418, ECO:0000269|PubMed:25301072}.; Involved in resistance response to the pathogenic oomycetes Phytophthora infestans and Phytophthora capsici. {ECO:0000269|PubMed:23980842, ECO:0000269|PubMed:25083911, ECO:0000269|PubMed:26795144}.

蛋白质序列

>L-type lectin-domain containing receptor kinase I.9 (LecRK-I.9) (LecRK79) (EC 2.7.11.1) (Protein DOES NOT RESPOND TO NUCLEOTIDES 1) (Purinoreceptor kinase 1)

MARWLLQILIISSLHLSSVSSQQETSFVYESFLDRQNLYLDKSAIVLPSGLLQLTNASEHQMGHAFHKKPIEFSSSGPLSFSTHFVCALVPKPGFEGGHGIVFVLSPSMDFTHAESTRYLGIFNASTNGSSSYHVLAVELDTIWNPDFKDIDHNHVGIDVNSPISVAIASASYYSDMKGSNESINLLSGNPIQVWVDYEGTLLNVSVAPLEVQKPTRPLLSHPINLTELFPNRSSLFAGFSAATGTAISDQYILWWSFSIDRGSLQRLDISKLPEVPHPRAPHKKVSTLIILLPVCLAILVLAVLAGLYFRRRRKYSEVSETWEKEFDAHRFSYRSLFKATKGFSKDEFLGKGGFGEVYRGNLPQGREIAVKRVSHNGDEGVKQFVAEVVSMRCLKHRNLVPLFGYCRRKRELLLVSEYMPNGSLDEHLFDDQKPVLSWSQRLVVVKGIASALWYLHTGADQVVLHRDVKASNIMLDAEFHGRLGDFGMARFHEHGGNAATTAAVGTVGYMAPELITMGASTGTDVYAFGVFMLEVTCGRRPVEPQLQVEKRHMIKWVCECWKKDSLLDATDPRLGGKFVAEEVEMVMKLGLLCSNIVPESRPTMEQVVLYLNKNLPLPDFSPYTLGIGTFAPVLVDASSLVVSSASWSLSGPSMSSSSPNHSPYAWQSTDQPWGQTIDTKNSLHIVAEPEKPSPAVKMVTLPAEDPQSNHSSISSQRVQPVKREKRRLHQILVAFPWINKQYFKLGLPKHIVHVSLFFFLQLARL

关键词

Alternative splicing;ATP-binding;Cell membrane;Glycoprotein;Kinase;Lectin;Membrane;Nucleotide-binding;Plant defense;Receptor;Reference proteome;Serine/threonine-protein kinase;Signal;Stress response;Transferase;Transmembrane;Transmembrane helix


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At5g60300(LECRK19) 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=At5g60300&sub_type=gene&SEARCH_EXACT=4&SEARCH_CONTAINS=1
NCBI网站：https://www.ncbi.nlm.nih.gov/gene/?term=836152
Uniprot 网站：https://www.uniprot.org/uniprotkb//entry

相关文献 (NCBI PubMed ID)

PMID: 27862469; 14593172; 16361528; 19773388; 21483488; 25301072; 25083911; 23980842; 24436418; 26795144