质粒编号
质粒名称
规格
价格
V003187
pSilent-1
5 μg (冻干粉)
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。
Based on established molecular biology principles (though not explicitly detailed in the provided search results), the pSilent-1 vector is a specialized plasmid designed for RNA interference (RNAi) studies. Its core functions include: Gene Silencing: Expresses short hairpin RNA (shRNA) to target and degrade specific mRNA, enabling targeted gene knockdown in research models. Controlled Expression: Utilizes an inducible promoter (e.g., Pol III promoters like U6 or H1) to regulate shRNA transcription, minimizing off-target effects. Selection & Screening: Incorporates antibiotic resistance markers (e.g., ampicillin or neomycin) for stable cell line selection post-transfection. In summary, pSilent-1 facilitates precise, reversible gene silencing to study gene function and validate therapeutic targets.
- 载体名称:
- pSilent-1
- 载体抗性:
- Ampicillin
- 载体长度:
- 7059 bp
- 载体类型:
- Ascomycete silencing vector
- 复制子:
- ori
- 载体来源:
- Nakayashiki H, Hanada S, Nguyen BQ, Kadotani N, Tosa Y, Mayama S.
- 启动子:
- trpC
- 感受态:
- stbl3
- 培养温度:
- 37℃
pSilent-1 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。
pSilent-1 载体序列
LOCUS Exported 7059 bp DNA circular SYN 14-JUL-2025
DEFINITION Ascomycete silencing vector pSilent-1, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7059)
AUTHORS Nakayashiki H, Hanada S, Nguyen BQ, Kadotani N, Tosa Y, Mayama S.
TITLE RNA silencing as a tool for exploring gene function in ascomycete
fungi
REFERENCE 2 (bases 1 to 7059)
AUTHORS De Schamphelaire W, Olbrechts A, Meert J, Verhelst K, Roggeman
Fonseca M, Vanhoucke M, Beyaert R.
TITLE BCCM/LMBP Plasmid collection
JOURNAL Unpublished
REFERENCE 3 (bases 1 to 7059)
AUTHORS De Schamphelaire W.
TITLE Direct Submission
JOURNAL Submitted (15-MAR-2017) BCCM/LMBP, Universiteit Gent,
Technologiepark 927, 9052, BELGIUM
REFERENCE 4 (bases 1 to 7059)
TITLE Direct Submission
REFERENCE 5 (bases 1 to 7059)
TITLE Direct Submission
REFERENCE 6 (bases 1 to 7059)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName:
"Unpublished"
COMMENT SGRef: number: 3; type: "Journal Article"; journalName: "Submitted
(15-MAR-2017) BCCM/LMBP, Universiteit Gent, Technologiepark 927,
9052, BELGIUM"
COMMENT SGRef: number: 4; type: "Journal Article"
COMMENT SGRef: number: 5; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..7059
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 24..128
/label=AmpR promoter
CDS 129..986
/label=AmpR
/note="beta-lactamase"
rep_origin 1160..1748
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind 2036..2057
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 2072..2102
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2110..2126
/label=lac operator
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
primer_bind 2134..2150
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
promoter 2171..2189
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 2234..2588
/label=trpC promoter
/note="promoter for Aspergillus nidulans trpC"
CDS 2593..3615
/label=HygR
/note="aminoglycoside phosphotransferase from E. coli"
regulatory 3639..4123
/label=trpC terminator
/note="trpC terminator"
/regulatory_class="terminator"
primer_bind 4132..4148
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5444..5460)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
intron 5491..5624
/note="CUT intron"
terminator 5829..6396
/label=trpC terminator
/note="transcription terminator from the Aspergillus
nidulans trpC gene"
promoter complement(6416..6434)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(6444..6460)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
rep_origin 6602..7057
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"