pCDNA3.1-EGFP①哺乳动物细胞绿色荧光表达载体质粒

pCDNA3.1-EGFP①哺乳动物细胞绿色荧光表达载体质粒
启动子: CMV promoter
复制子: pUC ori，F1 ori
终止子: BGH poly（A）signal
质粒分类: 哺乳系列质粒；哺乳表达质粒；pCDNA系列质粒
质粒大小: 6173bp
质粒标签: C-EGFP
原核抗性: 氨苄青霉素Amp（100μg/ml）
筛选标记: 新霉素Neo/G418
克隆菌株: DH5α等大肠杆菌
培养条件: 37℃，有氧，LB
表达宿主: 293T等哺乳细胞
培养条件: 37℃，5%CO2
诱导方式: 无需诱导
5'测序引物: pCDNA3.1-F（CTAGAGAACCCACTGCTTAC）
3'测序引物: pCDNA3.1-R（TAGAAGGCACAGTCGAGG）
质粒简介
pCDNA3.1-EGFP质粒是将EGFP基因克隆进入pCDNA3.1载体所得，设计用于哺乳动物宿主中的高水平稳定和瞬时表达。大多数哺乳动物细胞可以进行高水平的稳定和非复制性瞬时表达。人类巨细胞病毒立即早期（CMV）启动子，用于在广泛哺乳动物细胞中的高水平表达。在潜伏感染SV40或表达SV40大T抗原（例如COS-1，COS-7）的细胞系中的异常复制。
The GFP in pcDNA3.1-eGFP stands for green fluorescent protein. Deoxyribonucleic acids (DNAs) are typically made up of protein, so it makes sense for a special protein to make up a specific type of DNA add-gene such as the pcDNA3.1.although other means besides cloning would also be openly discussed. In this study, a detailed overview of how pcDNA-eGFP can be produced through cloning is done thanks to the use of secondary sources; a proper review of the pcDNA3.1-eGFP is first carried out to understand this compound. Afterward, the importance of a vector in the cloning process is looked at in detail and finally the cloning system is covered extensivel GFP proteins are generally composed of 238 amino acids with molecular masses of around 26.9 KD [1] and are typically used in the field of molecular biology as one of the many reporter genes. Basically, a reporter gene is a type of gene that researchers, especially in laboratory experiments, use to attach to a pre-specified sequence of the gene (oftentimes an experimental one) such as that of bacteria, plants, animals, and cell cultures. Some of the criteria that researchers use when selecting a reporter gene include, but may not be limited to, having easily identifiable and selectable markers and their ability to introduce changes that tend to be easily spotted under certain conditions. When conducting laboratory experiments in molecular biology, it would be important for researchers to know the different variables involved and the impact that they create on the experimental environment. Therefore, it makes sense to select reporter genes that possess these qualities such as the GFP [1]. In previously published studies involving the use of GFPs, including but not limited to pcDNA3.1, it has been common for researchers to introduce the GFP gene into cells using vector-based systems. In some cases, the researchers also used recombinant viruses (attaching the GFP to them). Being used as a reporter protein, the location of the target protein can be easily identified and expressed. However, in many of the laboratory experiments, the selection market of the GFPs used was not specific enough and there is often no selection market to normalize the transfection among other reactions.
质粒图谱

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