pCMV-MEKK1哺乳动物表达载体说明

pCMV-MEKK1

编号 载体名称

北京华越洋生物VECT6118 pCMV-­‐MEKK1

pCMV-­‐MEKK1载体基本信息

载体名称: pCMV-MEKK1

质粒类型: 哺乳动物表达载体；基因表达载体高拷贝/低拷贝: 高拷贝

克隆方法: 限制性内切酶，多克隆位点

启动子: CMV

载体大小: 5 kb

5' 测序引物及序列: --

3' 测序引物及序列: --

载体标签: 无

载体抗性: 卡那霉素

筛选标记: Neomycin

克隆菌株: TOP10F′, JM109 and INVaF′

宿主细胞（系）: 常规细胞系，如293、Hela等

备注: pCMV-MEKK1载体是蛋白激酶MEKK1的表达载体；CMV启动子驱动MEKK1基因的过表达；

MEKK1蛋白激酶是JNK/SAPK信号通路的应答激酶。

产品目录号: 631927

稳定性: 瞬表达或稳表达

组成型/诱导型: 组成型

病毒/非病毒: 非病毒

pCMV-­‐MEKK1载体质粒图谱和多克隆位点信息

pCMV-MEKK1载体使用说明

pCMV-MEKK1 is designed for the constitutive expression of MEK Kinase 1 (MEKK1) in mammalian cells. It can also be used to study the effects of a given stimulus on the JNK/SAPK pathway. In conjunction with Clontech’s In Vivo Kinase Assay Kits, pCMV-MEKK1 is an ideal positive control for studying a gene or molecule and its affect on kinase activation in the JNK/SAPK pathway.

MEKK1 mediates cellular responses to diverse stimuli, such as cytokines, growth factors, and other stressors via activation of JNK/SAPK signaling pathway. It has also been reported that MEKK1 is a possible regulator in signal transduction pathways involving apoptosis (1). To a lesser extent, MEKK1 can also activate the ERK pathway via phosphorylation of MEK (2, 3); however, it is more efficient in activating the JNK/SAPK than ERK (4, 5).

pCMV-MEKK1 contains a gene encoding MEKK1 (mouse) that is driven by the human cytomegalovirus(CMV) promoter. SV40 polyadenylation signals downstream of the MEKK1 gene direct proper processing of the 3’ end of the mRNA. The vector backbone also contains an SV40 origin for replication in mammalian cells expressing the SV40 T-antigen. A neomycin resistance cassette (Neor)—consisting of the SV40 early promoter, the neomycin/kanamycin resistance gene of Tn5, and polyadenylation signals from the Herpes simplex virus thymidine kinase (HSV TK) gene—allows stably transfected eukaryotic cells to be selected using G418(6). A bacterial promoter upstream of this cassette expresses kanamycin resistance in E. coli. The pCMV-MEKK1 backbone also provides a pUC origin of replication for propagation in E.coli and an f1 origin for single-stranded DNA production. The recombinant vector

can be transfected into mammalian cells using any standard method.

Propagation in E. coli:

Suitable host strains: DH5α, HB101, and other general purpose strains. Single-stranded DNA production requires

a host containing an F plasmid such as JM109 or XL1-Blue.

Selectable marker: plasmid confers resistance to kanamycin (50 μg/ml) to E. coli hosts.

E. coli replication origin: pUC

Copy number: ≈500

Plasmid incompatibility group: pMB1/ColE1

其他哺乳动物表达载体：

pCHO1.0 pBApo-CMV-Pur pOPRSVI

pcDNA3.1/His C pcDNA5/FRT/V5-His-TOPO pREP4

pcDNA3.1/His B pcDNA5/FRT/TO-TOPO pDual-GC

pcDNA3.1/His A pcDNA5/TO pBK-RSV

pIRESpuro3 pcDNA5/FRT/TO pBK-CMV

pIRES2-EGFP pcDNA5/FRT pBI-CMV4

pTT5 pFLAG-CMV2 pcDNA4/TO/Myc-His/LacZ pNFkB-DD-tdTomato pcDNA3.1/CT-GFP-TOPO pOPI3CAT

pBI-CMV5 pcDNA3.1/NT-GFP-TOPO pGene/V5-His B pSEAP2-Basic pOptiVEC-TOPO pSwitch

pSEAP2-Control pCMV-MEKK1 pCMVLacI

pBI-CMV3 pCMV-MEK1 pVgRxR

pBI-CMV2 pCMV-PKA pIND

pBI-CMV1 pcDNA6.2/nTC-Tag-DEST pTRE3G-Luc

pNFκB-MetLuc2-Reporter pcDNA6.2/cTC-Tag-DEST pTRE3G

pCRE-MetLuc2-Reporter pcDNA3.2/V5/GW/D-TOPO pTRE2-hygro

pAcGFP1-Actin pcDNA6.2/V5/GW/D-TOPO pTRE-Tight

pAcGFP1-N In-Fusion Ready pcDNA6.2/nGeneBLAzer-GW/D-TOPO pTK-hyg

pAcGFP1-C3 pcDNA6.2/C-YFP-DEST pTet-On

pAcGFP1-C pcDNA6.2/cGeneBLAzer-DEST pTet-Off

pAcGFP1-p53 pcDNA6/V5-His A pTet on advanced pAcGFP1-Mito pcDNA6/V5-His B pRevTRE

pAcGFP1-Mem pcDNA6/V5-His C pRevTet-On

pAcGFP1-Lam pcDNA6/myc-His C pRevTet-Off

pAcGFP1-Golgi pcDNA6/myc-His A pCMV-Tet3G

pAcGFP1-F pcDNA6/myc-His B pTRE2

pAcGFP1-Hyg-C1 pcDNA6.2/nGeneBLAzer-DEST pBD-NF-κB

pAsRed2-N1 pcDNA4/HisMax-TOPO pCMV-AD

ptdTomato-N1 pcDNA6.2/nLumio-DEST pCMV-BD

pCMV-tdTomato pcDNA6.2/cLumio-DEST pBIND-Id Control

pCRE-DD-tdTomato pcDNA4/myc-His C pBIND