pLVX-DsRed-Monomer-C1慢病毒红色荧光表达载体-BioVector NTCC质粒载体菌种细胞蛋白抗体基因保藏中心www.biovector.net

pLVX-DsRed-Monomer-C1慢病毒红色荧光表达载体

BioVector NTCC质粒载体菌种细胞蛋白抗体基因保藏中心www.biovector.net

基于HIV-1的慢病毒表达载体，pCMV-IE启动子，目的基因插入在DsRed-Monimer的C端，Amp抗性，puromycin细胞筛选标记。

Description

pLVX-DsRed-Monomer-C1 is an HIV-1-based, lentiviral expression vector that allows you

to express your gene of interest fused to DsRed-Monomer, a monomeric mutant of the

Discosoma sp. red fluorescent protein. Genes cloned into the multiple cloning site (MCS),

located at the C-terminal end of the DsRed-Monomer coding sequence, are expressed as

C-terminal fusions of the DsRed-Monomer protein. Expression of the fusion protein is driven

by the constitutively active human cytomegalovirus immediate early promoter (PCMV IE)

located just upstream of the DsRed-Monomer coding sequence. Lentiviral particles derived

from the vector allow the expression of DsRed-Monomer fusion proteins in virtually any cell

type, including primary cells. The unmodified vector expresses DsRed-Monomer, and may

be used to produce marker virus to optimize infection protocols.

pLVX-DsRed-Monomer-C1 contains all of the viral processing elements necessary for the

production of replication-incompetent lentivirus, as well as elements to improve viral

titer, transgene expression, and overall vector function. The woodchuck hepatitis virus

posttranscriptional regulatory element (WPRE) promotes RNA processing events and

enhances nuclear export of viral and transgene RNA (1), leading to increased viral titers from

packaging cells, and enhanced expression of your gene of interest in target cells. In addition,

the vector includes a Rev-response element (RRE), which further increases viral titers by

enhancing the transport of unspliced viral RNA out of the nucleus (2). Finally, pLVX-DsRed-

Monomer-C1 also contains a central polypurine tract (cPPT) element that increases nuclear

importation of the viral genome during target cell infection, resulting in improved vector

integration and more efficient transduction (3).

In addition to lentiviral elements, pLVX-DsRed-Monomer-C1 contains a puromycin resistance gene (Puror)

under the control of the murine phosphoglycerate kinase (PGK) promoter (PPGK) for the selection of stable

transductants. The vector also contains a pUC origin of replication and an E. coli ampicillin resistance gene

(Ampr) for propagation and selection in bacteria.

Use

pLVX-DsRed-Monomer-C1 constitutively expresses your gene of interest from PCMV IE when transduced into

target cells. Before the vector can be transduced into cells, however, it must be transfected into 293T packaging

cells with our Lenti-X™ HTX Packaging System. This packaging system allows

you to safely produce high titer, infectious, replication-incompetent, VSV-G pseudotyped lentiviral particles

that can infect a wide range of cell types, including non-dividing and primary cells (4).

Location of Features

• 5' LTR: 1–635

• PBS (primer binding site): 636–653

• Ψ (packaging signal): 685–822

• RRE (Rev-response element): 1303–1536

• cPPT(central polypurine tract): 2028–2151

• PCMV IE (human cytomegalovirus immediate early promoter): 2185–2788

• DsRed-Monomer (Discosoma sp. red fluorescent protein monomer): 2816–3490

• MCS (multiple cloning site): 3504–3556

• PPGK (phosphoglycerate kinase promoter): 3580–4088

• Puror (puromycin resistance gene ): 4109–4708

• WPRE (woodchuck posttranscriptional regulatory element): 4722–5313

• 3' LTR: 5517–6153

• pUC origin of replication: 6623–7293 (complementary)

• Ampr (ampicillin resistance gene; β-lactamase): 7438–8434 (complementary)

Selection of Stable Transfectants

• Selectable marker: plasmid confers resistance to puromycin.

Propagation in E. coli

• Suitable host strains: DH5α, DH10B and other general purpose strains.

• Selectable marker: plasmid confers resistance to ampicillin (100 μg/ml) in E. coli hosts.

• E. coli replication origin: pUC

• Copy number: high

Excitation and emission maxima of DsRed-Monomer

• Excitation maximum = 557 nm

• Emission maximum = 592 nm

Map图谱