■ 基本信息
启动子: | CMV |
复制子: | pUC |
质粒分类: | 哺乳细胞，蛋白过表达载体 |
质粒大小: | 5.3kb |
原核抗性: | Amp |
真核抗性: | G418 |
克隆菌株: | DH5a |
培养条件: | 37度 |
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: | CMV-F：CGCAAATGGGCGGTAGGCGTG |
3'测序引物: | 根据序列设计引物 |
■ 质粒属性
质粒宿主: | 哺乳细胞 |
质粒用途: | 蛋白表达 |
片段类型: | ORF |
片段物种: | 空载体 |
原核抗性: | Amp |
真核抗性: | G418
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荧光标记: |
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■ 质粒简介
pIRESneo (originally described as pCIN4 by Rees et al.; 1) contains the internal ribosome entry site (IRES) of the encephalomyocarditis virus (ECMV), which permits the translation of two open reading frames from one messenger RNA (2, 3). After selection with G418, nearly all surviving colonies will stably express the gene of interest, thus decreasing the need to screen large numbers of colonies to find functional clones. The expression cassette of pIRESneo contains the human cytomegalovirus (CMV) major immediate early promoter/enhancer followed by a multiple cloning site (MCS), a synthetic intron known to enhance the stability of the mRNA (4), the ECMV IRES followed by the neomycin phosphotransferase (NPT II) gene, and the polyadenylation signal of the bovine growth hormone. Ribosomes can enter the bicistronic mRNA either at the 5' end to translate the gene of interest or at the ECMV IRES to translate the antibiotic resistance marker.
质粒只保证关键序列正确，不保证表达效果。
■ 质粒图谱
