pECFP-C1质粒

基本信息
启动子: CMV
复制子: pUC ori,f1 ori
终止子: SV40 poly(A) signal
质粒分类: 哺乳细胞，荧光蛋白报告载体
质粒大小: 4731bp
原核抗性: Kan 筛选标记: Neo
克隆菌株: DH5α
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: CMV-F：CGCAAATGGGCGGTAGGCGTG
3'测序引物: Sv40-polyA-R：GAAATTTGTGATGCTATTGC
质粒简介
pECFP-C1 encodes an enhanced cyan fluorescent variant of the Aequorea victoria green fluorescent protein gene (GFP). The ECFP gene contains six amino acid substitutions. The Tyr-66 to Trp substitution gives ECFP fluorescence excitation (major peak at 433 nm and a minor peak at 453 nm) and emission (major peak at 475 nm and a minor peak at 501 nm) similar to other cyan emission variants (1–3). The other five substitutions (Phe-64 to Leu; Ser-65 to Thr; Asn-146 to Ile; Met-153 to Thr; and Val-163 to Ala) enhance the brightness and solubility of the protein, primarily due to improved protein-folding properties and efficiency of chromophore formation (2, 4, 5).