■ 基本信息
別名: | 和2酶切位點(diǎn)不一樣 |
啟動(dòng)子: | CMV |
復制子: | pUC |
終止子: | BGH poly(A)signal |
質(zhì)粒分類(lèi): | 哺乳系列質(zhì)粒;哺乳表達質(zhì)粒;pCDNA系列質(zhì)粒 |
質(zhì)粒大小: | 6173bp |
質(zhì)粒標簽: | C-EGFP |
原核抗性: | Amp |
真核抗性: | G418 |
克隆菌株: | DH5a |
培養條件: | 37℃,5%CO2 |
表達宿主: | 293T等哺乳細胞 |
誘導方式: | 無(wú)需誘導 |
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: | 哺乳細胞 |
質(zhì)粒用途: | 蛋白表達 |
片段類(lèi)型: | ORF
|
片段物種: |
|
原核抗性: | Amp |
真核抗性: | G418
|
熒光標記: | 綠色
|
■ 質(zhì)粒簡(jiǎn)介
pCDNA3.1-EGFP質(zhì)粒是將EGFP基因克隆進(jìn)入pCDNA3.1載體所得,設計用于哺乳動(dòng)物宿主中的高水平穩定和瞬時(shí)表達。大多數哺乳動(dòng)物細胞可以進(jìn)行高水平的穩定和非復制性瞬時(shí)表達。人類(lèi)巨細胞病毒立即早期(CMV)啟動(dòng)子,用于在廣泛哺乳動(dòng)物細胞中的高水平表達。在潛伏感染SV40或表達SV40大T抗原(例如COS-1,COS-7)的細胞系中的異常復制。
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: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