Note: The 1.3xWT HBV - Luciferase vector is an important tool for studying hepatitis B virus (HBV) gene expression.
It is constructed by replacing specific sequences in the HBV genome with the luciferase gene, enabling the measurement of HBV transcriptional activity through luciferase activity. Its key characteristics include a specific start codon from the Core ORF and regulation by the pgRNA promoter, ensuring accurate reflection of HBV gene expression changes. The main advantages are its high specificity and sensitivity in detecting HBV transcriptional activity and its usefulness in studying the roles of different regulatory elements in the HBV genome.
It should be chosen when investigating HBV gene expression regulation, particularly when analyzing the effects of various factors on HBV transcription or when exploring the interactions between different regulatory elements within the HBV genome.
基本信息
- 载体名称:
- 1.3xWT HBV-Luciferase
- 载体抗性:
- Ampicillin
- 载体长度:
- 6656 bp
- 载体类型:
- Mammalian Expression, Luciferase
- 复制子:
- ori
- 启动子:
- SP6
- 感受态:
- DH10B
- 培养温度:
- 37℃
产品信息
质粒编号:V000707
质粒名称:1.3xWT HBV-Luciferase
规格:5 μg (冻干粉)
下载资源
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。
确保质粒的关键元件正确，但是我们并不能保证实验效果。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。
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。
1.3xWT HBV-Luciferase 质粒 (编号: V000707)序列
LOCUS V000707 6656 bp DNA circular SYN 13-MAY-2021
DEFINITION Exported.
ACCESSION V000707
VERSION V000707
KEYWORDS 1.3xWT HBV-Luciferase
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 6656)
AUTHORS Doitsh G, Shaul Y
TITLE Enhancer I predominance in hepatitis B virus gene expression.
JOURNAL Mol Cell Biol. 2004 Feb;24(4):1799-808.
PUBMED 14749394
REFERENCE 2 (bases 1 to 6656)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 6656)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Mol Cell
Biol."; date: "2004-02"; volume: "24(4)"; pages: "1799-808"
SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..6656
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS 355..816
/gene="X"
/label="Protein X"
/note="Protein X from Hepatitis B virus genotype A2 subtype
adw2 (strain Rutter 1979). Accession#: P69713"
CDS 969..2618
/label="luciferase"
/note="firefly luciferase"
CDS 3317..3778
/gene="X"
/label="Protein X"
/note="Protein X from Hepatitis B virus genotype A2 subtype
adw2 (strain Rutter 1979). Accession#: P69713"
promoter complement(3981..3999)
/label="SP6 promoter"
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(4017..4033)
/label="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(4041..4057)
/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)."
promoter complement(4065..4095)
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind complement(4110..4131)
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4248..4265)
/label="L4440"
/note="L4440 vector, forward primer"
rep_origin complement(4419..5007)
/direction=LEFT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(5181..6038)
/label="AmpR"
/note="beta-lactamase"
promoter complement(6039..6143)
/label="AmpR promoter"
primer_bind 6211..6229
/label="pBRforEco"
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(6267..6289)
/label="pGEX 3'"
/note="pGEX vectors, reverse primer"
primer_bind 6389..6408
/label="pRS-marker"
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 6617..6633
/label="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
promoter 6640..6656
/label="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"