我们所展示的质粒图谱主要是从文献和开放数据库中收集而来，主要是为了方便研究工作，其中一小部分质粒进行了质量控制，可供科学家使用。
所有的产品都严格仅供科学研究使用，不能应用于临床试验，包括人体摄入、注射或外用的药理学用途。
确保质粒的关键元件正确，但是我们并不能保证实验结果。页面展示的图谱序列为理论序列，可能与测序结果不一致，请自行比对后确定是否满足要求。(如果测过序，本页面一般会提供下载)
开放数据库中的大多数载体序列都没有被完全测序。如果实际序列与参考序列的相似度超过99%，则将其视为正确。
由于科学研究是在探索未知，具有很大的不确定性，在任何情况下，我们都不承担超出质粒本身的额外经济损失或责任。
Modified Tobacco Rattle Virus RNA 2; used for virus induced gene silencing in plants along with pYL192 (TRV RNA1)
- 载体名称:
- pYL156 (TRV RNA2)
- 载体抗性:
- Kanamycin
- 载体长度:
- 11003 bp
- 载体类型:
- Plant Vectors
- 载体来源:
- Savithramma Dinesh-Kumar
- 拷贝数:
- High copy number
- 感受态:
- DH10B
- 培养温度:
- 37℃
pYL156 (TRV RNA2) 载体图谱
质粒操作方法
1. 发货形式：质粒干粉(常温运输，存于-20度，请务必先转化提质粒后使用)
2. 收到质粒干粉后请先5000rpm离心1min，再加入20μl ddH2O溶解质粒;（质粒复测的浓度有时候与标称值差距较大，这可能是因为冻干质粒在管中的位置、复溶效率、测量偏差以及管壁的吸附导致，因此建议先转化提质粒后再使用）
3. 取1支100μl 感受态于冰上解冻10min，加入2μl质粒，再冰浴30min后，42℃热激60s，不要搅动，再冰浴2min；
4. 加入900μl无抗的LB液体培养基，180rpm震荡37℃培养45min (30℃培养1-1.5小时);
5. 6000rpm离心5min，仅留100μl上清液重悬细菌沉淀，并涂布至目标质粒抗性的LB平板上;
6. 将平板倒置37℃培养14h，如果要求是30℃则培养20h; (菌落过多则将质粒稀释后再转化；没有菌落则加入10μl质粒转化；建议不要直接转表达感受态， 要先转克隆感受态，重提质粒后再导入表达感受态);
7. 挑取单菌落至LB液体培养基中，加入对应抗生素，220rpm震荡培养14h，根据实验需要和质粒提取试剂盒说明书提取质粒。
pYL156 (TRV RNA2) 载体序列
LOCUS Exported 11003 bp DNA circular SYN 06-DEC-2024
DEFINITION Modified Tobacco Rattle Virus RNA 2; used for virus induced gene
silencing in plants along with pYL192 (TRV RNA1).
ACCESSION .
VERSION .
KEYWORDS pYL156 (TRV RNA2)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 11003)
AUTHORS Liu Y, Schiff M, Marathe R, Dinesh-Kumar SP
TITLE Tobacco Rar1, EDS1 and NPR1/NIM1 like genes are required for
N-mediated resistance to tobacco mosaic virus.
JOURNAL Plant J. 2002 May;30(4):415-29. doi:
10.1046/j.1365-313x.2002.01297.x.
PUBMED 12028572
REFERENCE 2 (bases 1 to 11003)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; doi:
"10.1046/j.1365-313x.2002.01297"; journalName: "Plant J"; date:
"2002-05"; volume: "30"; issue: "4"; pages: "415-29"
FEATURES Location/Qualifiers
source 1..11003
/mol_type="other DNA"
/organism="synthetic DNA construct"
source 770..2408
/mol_type="other DNA"
/note="Tobacco rattle virus RNA2-based T-DNA vector;
synonym: pYL156"
/db_xref="taxon:188057"
/organism="Tobacco rattle virus RNA2-based VIGS vector
pTRV2"
source 2478..2872
/mol_type="other DNA"
/note="Tobacco rattle virus RNA2-based T-DNA vector;
synonym: pYL156"
/db_xref="taxon:188057"
/organism="Tobacco rattle virus RNA2-based VIGS vector
pTRV2"
promoter 427..769
/label=CaMV 35S promoter
/note="strong constitutive promoter from cauliflower mosaic
virus"
primer_bind 715..735
/label=35S promoter
/note="CaMV 35S promoter, forward primer"
misc_feature 770..2408
/label=TRV strain ppk20 RNA2 5'-sequence
/note="TRV strain ppk20 RNA2 5'-sequence"
misc_feature 2478..2872
/label=TRV strain ppk20 RNA2 3'-sequence
/note="TRV strain ppk20 RNA2 3'-sequence"
CDS 2933..2950
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
terminator 2985..3237
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
misc_feature 3259..3283
/label=RB T-DNA repeat
/note="right border repeat from nopaline C58 T-DNA"
CDS 4584..5213
/codon_start=1
/product="stability protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
/label=pVS1 StaA
/translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
CDS 5642..6715
/codon_start=1
/product="replication protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
/label=pVS1 RepA
/translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
LVMRYRNLIEGEASAGS"
rep_origin 6781..6975
/label=pVS1 oriV
/note="origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
primer_bind complement(7211..7233)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
misc_feature 7319..7459
/label=bom
/note="basis of mobility region from pBR322"
primer_bind 7374..7393
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind complement(7474..7491)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(7645..8233)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(7725..7744)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS 8404..9612
/codon_start=1
/label=IS4-like element ISVsa5 family transposase
/translation="MCELDILHDSLYQFCPELHLKRLNSLTLACHALLDCKTLTLTELG
RNLPTKARTKHNIKRIDRLLGNRHLHKERLAVYRWHASFICSGNTMPIVLVDWSDIREQ
KRLMVLRASVALHGRSVTLYEKAFPLSEQCSKKAHDQFLADLASILPSNTTPLIVSDAG
FKVPWYKSVEKLGWYWLSRVRGKVQYADLGAENWKPISNLHDMSSSHSKTLGYKRLTKS
NPISCQILLYKSRSKGRKNQRSTRTHCHHPSPKIYSASAKEPWILATNLPVEIRTPKQL
VNIYSKRMQIEETFRDLKSPAYGLGLRHSRTSSSERFDIMLLIALMLQLTCWLAGVHAQ
KQGWDKHFQANTVRNRNVLSTVRLGMEVLRHSGYTITREDSLVAATLLTQNLFTHGYVL
GKL"
CDS complement(9658..10452)
/codon_start=1
/gene="aphA-3"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin"
/translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
misc_feature 10877..10901
/label=LB T-DNA repeat
/note="left border repeat from nopaline C58 T-DNA"