质粒类型: | 慢病毒载体 |
---|---|
高拷贝/低拷贝: | 高拷贝 |
启动子: | EF1α/ EF1a |
克隆方法: | 多克隆位点,限制性内切酶 |
载体大小: | 9519 bp |
5' 测序引物及序列: | EF1a Forward: TCAAGCCTCAGACAGTGGTTC |
载体标签: | C-AcGFP1 |
载体抗性: | Ampicillin (氨苄青霉素) |
筛选标记: | Puromycin (嘌呤霉素) |
备注: | 载体能够表达C端AcGFP1荧光蛋白 |
产品编号 | 产品名称 | 规格 | 价格 |
---|---|---|---|
QC2008 | pLVX-EF1α-AcGFP1-N1 |
5ug质粒 |
¥1500.00 |
pLVX-EF1α-AcGFP1-N1 is an HIV-1-based, lentiviral expression vector designed to constitutively express a protein of interest fused to the N-terminus of AcGFP1, a green fluorescent protein derived from Aequorea coerulescens. The excitation and emission maxima of native AcGFP1 are 475 nm and 505 nm, respectively. Stable, constitutive expression of the fusion protein is driven by the EF1α promoter (PEF1α), which continues to be constitutively active even after the vector integrates into the host cell genome (1).
pLVX-EF1α-AcGFP1-N1 contains all of the viral processing elements necessary for the production of replicationincompetent 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 RNA (2), leading to increased viral titers from packaging 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 (3). Finally, pLVX-EF1α-AcGFP1-N1 also contains a central polypurine tract/central termination sequence element (cPPT/CTS). During target cell infection, this element creates a central DNA flap that increases nuclear import of the viral genome, resulting in improved vector integration and more efficient transduction (4).
In addition to lentiviral elements, pLVX-EF1α-AcGFP-N1 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.