■ 基本信息
別名: | 和2酶切位點(diǎn)不一樣 |
啟動子: | CMV |
復(fù)制子: | pUC |
終止子: | BGH poly(A)signal |
質(zhì)粒分類: | 哺乳系列質(zhì)粒;哺乳表達(dá)質(zhì)粒;pCDNA系列質(zhì)粒 |
質(zhì)粒大小: | 6173bp |
質(zhì)粒標(biāo)簽: | C-EGFP |
原核抗性: | Amp |
真核抗性: | G418 |
克隆菌株: | DH5a |
培養(yǎng)條件: | 37℃,5%CO2 |
表達(dá)宿主: | 293T等哺乳細(xì)胞 |
誘導(dǎo)方式: | 無需誘導(dǎo) |
5'測序引物:
| pCDNA3.1-F(CTAGAGAACCCACTGCTTAC)
|
3'測序引物: | pCDNA3.1-R(TAGAAGGCACAGTCGAGG) |
■ 質(zhì)粒屬性
質(zhì)粒宿主: | 哺乳細(xì)胞 |
質(zhì)粒用途: | 蛋白表達(dá) |
片段類型: | ORF
|
片段物種: |
|
原核抗性: | Amp |
真核抗性: | G418
|
熒光標(biāo)記: | 綠色
|
■ 質(zhì)粒簡介
pCDNA3.1-EGFP質(zhì)粒是將EGFP基因克隆進(jìn)入pCDNA3.1載體所得,設(shè)計用于哺乳動物宿主中的高水平穩(wěn)定和瞬時表達(dá)。大多數(shù)哺乳動物細(xì)胞可以進(jìn)行高水平的穩(wěn)定和非復(fù)制性瞬時表達(dá)。人類巨細(xì)胞病毒立即早期(CMV)啟動子,用于在廣泛哺乳動物細(xì)胞中的高水平表達(dá)。在潛伏感染SV40或表達(dá)SV40大T抗原(例如COS-1,COS-7)的細(xì)胞系中的異常復(fù)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.
本質(zhì)粒的參考文獻(xiàn):A novel long non-coding RNA, lncKBTBD10, involved in bovine skeletal muscle myogenesis.
質(zhì)粒只保證關(guān)鍵序列正確,不保證表達(dá)效果。
■ 質(zhì)粒圖譜

■ 質(zhì)粒序列
質(zhì)粒序列請下載:
ZK430pCDNA3.1-EGFP①哺乳熒光質(zhì)粒.txt