pCMV-MEK1
pCMV-MEK1
编号 | 载体名称 |
北京华越洋生物VECT6168 | pCMV-MEK1 |
pCMV-MEK1载体基本信息
载体名称: | pCMV-MEK1 |
质粒类型: | 哺乳动物表达载体；基因载体 |
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: | HB101, DH5alpha |
宿主细胞（系）: | 常规细胞系，如293、Hela等 |
备注: | pCMV-MEK1载体是蛋白激酶MEK1的表达载体；CMV启动子驱动基因的过表达。 |
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-MEK1载体质粒图谱和多克隆位点信息
pCMV-MEK1载体简介
pCMV-MEK1 is designed for the constitutive expression of MAPK/ERK kinase 1 (MEK 1) in mammalian cells. It can also be used to study the effects of a given stimulus on the MAPK signaling pathway. In conjunction with Clontech's In Vivo Kinase Assay Kits, pCMV-MEK1 is an ideal positive control for studying a gene or molecule and its affect on kinase activation in the MAPK pathway. MEK1, a Raf isoform, phosphorylates and activates ERK-1 and ERK-2 (1–3). Studies show that ERK1/2 are activated by mitogens in most cell types, linking their involvement in mitogenic signaling (2). ERKs may also be linked to cell transformation, differentiation, and apoptosis in some cell types (ibid.).
pCMV-MEK1 contains the gene encoding MEK1 (human), which is controlled by the human cytomegalovirus (CMV) promoter. SV40 polyadenylation signals downstream of MEK1 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 (4). A bacterial promoter upstream of this cassette expresses kanamycin resistance in E. coli. The pCMV-MEK1 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
pCMV-MEK1其他哺乳动物表达载体：
