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pLenti-Bi-cistronic质粒

货号：kl-zl-0723-01

货号	产品名称	规格	库存	价格	数量	购买
kl-zl-0723-01	pLenti-Bi-cistronic质粒	NA		立即咨询	+ -

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pLenti-Bi-cistronic质粒货号：kl-zl-0723 规格：20ul

启动子：	RSV ,PGK
复制子：	M13 ori,ori ,SV40 ori
终止子：	SV40 poly(A) signal
质粒分类：	病毒系列，慢病毒双表达载体
质粒大小：	8292bp
原核抗性：	Kan
筛选标记：	Puro
克隆菌株：	Stbl3
培养条件：	37℃，有氧 LB
表达宿主：	哺乳细胞
诱导方式：	无须诱导，瞬时表达
5'测序引物：	根据序列设计引物
3'测序引物：	根据序列设计引物

质粒简介：
This study was aimed to construct, package and purify the recombinant lentivirus vector carrying the firefly luciferase gene (FLUC) and red fluorescent protein gene (RFP) and to transfect the recombinant lentivirus into HeLa cells, so as to observe the expression levels of these two genes. The FLUC and RFP genes were amplified by RT-PCR and inserted in the lentiviral expression vector (pLenti-Bi-cistronic) to construct the lentiviral vector pLenti-FLUC-RFP. The viral particles were generated by cotransfection of 293T cells with pLenti-FLUC-RFP and three packaging vectors, and the virus titer was determined by calculating the percentage of RFP positive cells. After transfection of pLenti-FLUC-RFP into HeLa cells, the expression of RFP was observed by fluorescent microscopy, and the activity of FLUC was determined by luciferase reporter gene assay kit. The results showed that the inserting orientation of the RFP and FLUC genes in the lentiviral vector pLenti-FLUC-RFP were verified by restriction analysis. Targeted RFP and FLUC sequences were confirmed by DNA sequencing. The final titer obtained was 1×10TU/ml. The expressions of RFP and FLUC were observed in the transfected HeLa cells. It is concluded that the pLenti-III-FLUC-RFP recombinant lentivirus vector carrying RFP gene and FLUC gene with high viral titer is constructed and packaged successfully, and provides experimental basis for studying dynamic distribution of mesenchymal stem cells in vivo.
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