pGL4.43載體(ti) 基本信息

　　出品公司:Promega

　　載體(ti) 名稱:pGL4.43 ; pGL4.43[luc2P/XRE/Hygro]

　　質粒類型:信號通路報告載體(ti) ;哺乳動物載體(ti) ;螢火蟲熒光素酶報告載體(ti)

　　高拷貝/低拷貝:--

　　克隆方法:限製性內(nei) 切酶，多克隆位點

　　啟動子:Minimal Promotor

　　載體(ti) 大小:6067 bp

　　5' 測序引物及序列:--

　　3' 測序引物及序列:--

　　載體(ti) 標簽:luc2P

　　載體(ti) 抗性:氨苄青黴素

　　篩選標記:潮黴素(Hygromycin)

　　克隆菌株:TOP10等常規菌株

　　宿主細胞(係):HEK293等

　　備注:pGL4.43[luc2P/XRE/Hygro]載體(ti) 是信號通路報告載體(ti) ;含異己物應答元件XRE;含螢火蟲熒光素酶報告基因luc2P。

　　產(chan) 品目錄號:E3751

　　穩定性:穩表達

　　組成型/誘導型:組成型

　　病毒/非病毒:非病毒

　　pGL4.43載體(ti) 質粒圖譜和多克隆位點信息

　　pGL4.43載體(ti) 簡介

　　The pGL4.43[luc2P/XRE/Hygro] Vector contains three copies of a xenobiotic response element (XRE) that drives transcription of the luciferase reporter gene luc2P (Photinus pyralis). luc2P is a synthetically-derived luciferase sequence with humanized codon optimization that is designed for high expression and reduced anomalous transcription. The luc2P gene contains hPEST, a protein destabilization sequence, which allows luc2P protein levels to respond more quickly than those of luc2 to induction of transcription. The vector backbone contains an ampicillin resistance gene to allow selection in E. coli and a gene for hygromycin resistance to allow selection of stably transfected mammalian cell lines.

　　Example Protocol

　　In this example protocol, the pGL4.43[luc2P/XRE/Hygro] Vector is used to measure

　　activation of the XRE in HepG2 cells upon treatment with TCDD. The pGL4.75 Vector

　　(encoding Renilla luciferase) is used as a normalization control. In designing such

　　experiments, it is important that the chosen cell type can be transfected efficiently and

　　that it expresses the proper components of the signaling pathway of interest in order to

　　generate the biological response. Protocol optimization may be required for your

　　particular cell type and assay conditions.

　　實驗材料

　　DMEM (Life Technologies Cat.# 11995)

　　Complete medium [DMEM supplemented with 10% fetal bovine serum (DMEM/FBS;

　　Life Technologies Cat.# 16000] and 1X NEAA [Life Technologies Cat.# 11140])

　　Dulbecco’s PBS (DPBS; Life Technologies Cat. # 14190)

　　0.05% Tryspin-EDTA (Life Technologies Cat.# 25300)

　　Charcoal-stripped FBS (Life Technologies Cat.# 126776-011)

　　Opti-MEM I (Life Technologies Cat.# 31985)

　　FuGENE HD Transfection Reagent (Cat.# E2311)

　　TCDD (2.3.7.8-Tetrachlorodibenzo-p-dioxin; AccuStandard Cat.# D-404N)

　　DMSO (Sigma Cat.# D2650)

　　Dual-Glo Luciferase Assay System (Cat.# E2940)

　　HepG2 cells

　　pGL4.75[hRluc/CMV] Vector (Cat.# E6931)

　　質粒載體(ti) 網主要對載體(ti) 質粒類目下產(chan) 品展示,不僅(jin) 提供質粒、菌種、細胞及培養(yang) 基等相關(guan) 產(chan) 品訂購,同時可以為(wei) 客戶提供質粒構建、載體(ti) 設計及菌種細胞鑒定,還對包括ATCC、JCM、KCTC等進口微生物資源提供訂 購及保藏服務。