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- 详细信息
- 文献和实验
- 技术资料
- 保存条件:
常温
- 保质期:
三年
- 英文名:
pcDNA3.1(+)
- 库存:
60
- 供应商:
信裕生物
- 规格:
5ug质粒
基本信息
| 质粒类型: | 哺乳动物表达载体 |
|---|---|
| 启动子: | CMV |
| 表达水平: | 高 |
| 克隆方法: | 多克隆位点,限制性内切酶 |
| 载体大小: | 5428 bp (查看载体序列) |
| 5' 测序引物: | pcDNA3.1-F:CTAGAGAACCCACTGCTTAC |
| 3' 测序引物: | pcDNA3.1-R (BGH-R):TAGAAGGCACAGTCGAGG |
| 载体抗性: | Ampicillin (氨苄青霉素) |
| 筛选标记: | Neomycin / G418 |
订购信息
| 产品编号 | 产品名称 | 规格 | 价格 |
|---|---|---|---|
| XY1001 | pcDNA3.1(+) | 5ug质粒 |
¥1000.00 |
质粒图谱
载体描述
pcDNA3.1(+) and pcDNA3.1(-) are 5.4 kb vectors derived from pcDNA3 and designed for high-level stable and transient expression in mammalian hosts. High-level stable and non-replicative transient expression can be carried out in most mammalian cells. The vectors contain the following elements:
- Human cytomegalovirus immediate-early (CMV) promoter for high-level expression in a wide range of mammalian cells
- Multiple cloning sites in the forward (+) and reverse (-) orientations to facilitate cloning
- Neomycin resistance gene for selection of stable cell lines
- Episomal replication in cells lines that are latently infected with SV40 or that express the SV40 large T antigen (e.g. COS-1, COS-7). The control plasmid, pcDNA3.1/CAT, is included for use as a positive control for transfection and expression in the cell line of choice.
This pcDNA™3.1(+)vector is designed for high-level, constitutive expression in a variety of mammalian cell lines. It contains a Geneticin® selectable marker and a forward-orientation multiple cloning site.
The pcDNA™3.1 Expression Vector Family
Three untagged versions of pcDNA™3.1 (available separately), each with a different selectable marker (Geneticin®, Zeocin™, or Hygromycin), are for use alone or in co-transfections. All three vectors offer the following features:
• Cytomegalovirus (CMV) enhancer-promoter for high-level expression
• Large multiple cloning site in either forward (+) or reverse (-) orientations
• Bovine Growth Hormone (BGH) polyadenylation signal and transcription termination sequence for enhanced mRNA stability
• SV40 origin for episomal replication and simple vector rescue in cell lines expressing the large T antigen (i.e., COS-1 and COS-7)
• Ampicillin resistance gene and pUC origin for selection and maintenance in E. coli
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文献和实验wangch84 各位高手,兄弟最近忙于质粒构建,意图将一段长约4000 bp的片段,通过BamH1和EcoR1两个酶切位点连接到pcDNA3.1+载体上。情况如下: 1. 通过LA Taq酶扩增,并纯化回收目的片段(引物中带有酶切位点,保护性碱基=3,上游带有Kozak序列);由于回收效率很高,所以在进行下一步双酶切时只加了12 μl的目的片段,H2O 20 μl,10 X K BUFFER 4 μl,两种酶各2 μl,37℃作用过夜(12-14 h
1.常用载体分类:2,常用载体元件介绍:(1) 启动子:启动子是位于结构基因5'端上游的DNA序列,能活化RNA聚合酶,使之与模板DNA准确的结合并具有转录起始的特异性。启动子主要分为广谱启动子和特异性启动子。广谱启动子为在各种组织、细胞上广泛表达的启动子;特异性启动子为在特异性组织或者细胞类型上表达的启动子。常用广谱启动子有CAG,CMV,EF1a,PGK等,还包含启动RNA的III型启动子U6和H1启动子;常用特异性启动子如成熟神经元特异性启动子hSyn,投射神经元特异性启动
恰当的选择固相载体和连接分子是固相反应的重要组成部分. 固相载体和连接分子都应该在多种遗传反应过程期间是保持稳定, 而且产品的切除应该是在温和的条件下实现. 本章将介绍各种不同的载体和连接分子. 如果反应后可以用过滤的方法去除, 从理论上讲, 任何材料都可用作固相载体. 聚乙烯, 聚苯乙烯-外涂聚乙烯薄层, 平滑的纸或棉花等都可以. 最常用的载体是: 聚苯乙烯(PS), 聚乙二醇(PEG), Tentagel (TG), 和PEGA (聚乙二醇-聚丙
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