pEYFP-Tub荧光蛋白报告载体

Read More
Thumbnail image

载体订购信息

产品编号
GS-1127
产品名称
pEYFP-Tub 荧光蛋白报告载体
市场价格
￥ 1000
沟通洽谈

载体基本信息

载体名称:
pEYFP-Tub
载体类型:
荧光蛋白报告载体
克隆方法:
/
拷贝类型:
/
载体抗性:
Kanamycin (卡那霉素)
筛选标记:
Neomycin（新霉素）
载体大小:
6045 bp 点击查看完整序列
启动子:
CMV
测序引物5’:
/
测序引物3’:
/
报告基因:
/

载体图谱展示

载体图谱

载体说明

pEYFP-Tub encodes a fusion protein consisting of enhanced yellow fluorescent protein (EYFP) and human a-tubulin (1, 2). EYFP is an enhanced yellow-green variant of the Aequorea victoria green fluorescent protein (GFP; 3–5). The EYFP gene contains the four amino acid substitutions previously published as GFP-10C (6): Ser-65 to Gly; Val-68 to Leu; Ser-72 to Ala; and and Thr-203 to Tyr. It also contains the Phe-64 to Leu mutation of GFPmut1 (7). The fluorescence excitation maximum of EYFP is 513 nm; the emission spectrum has a peak at 527 nm (in the yellow-green region). When excited at 513 nm, the Em of EYFP is 36,500 cm–1M–1 and the fluorescence quantum yield is 0.63 (6), resulting in a bright fluorescent signal. The fluorescence observed is roughly equivalent to that from EGFP.

In addition to the chromophore mutations, pEYFP-Tub contains more than 190 silent base changes that correspond to human codon-usage preferences (8). SV40 polyadenylation signals downstream of the EYFP-Tub fusion direct proper processing of the 3' end of the mRNA. The vector backbone also contains an SV40 origin for replication in mammalian cells expressing the SV40 T-antigen. A neomycin resistance cassette (Neor) consisting of the SV40 early promoter, the neomycin/ kanamycin resistance gene of Tn5, and polyadenylation signals from the herpes simplex virus thymidine kinase (HSV-TK) gene allows stably transfected eukaryotic cells to be selected using G418 (9). A bacterial promoter upstream of this cassette drives expression of the gene encoding kanamycin resistance in E. coli. The pEYFP-Tub backbone also provides a pUC origin of replication for propagation in E. coli and an f1 origin for single-stranded DNA production.

载体应用

The pEYFP-Tub Vector is designed for expressing the EYFP-tubulin fusion protein in mammalian cells. pEYFP-Tub can be introduced into mammalian cells using any standard transfection method. If required, stable transformants can be selected using G418 (9). The protein incorporates directly into microtubules and thereby allows the observation of microtubules in living or fixed cells by fluorescence microscopy (10). pEYFP-Tub is not sold as a cloning vector; however, there are unique restriction sites at the 5' end of EFYP, at the EYFP-Tub fusion junction, and at the 3' end of the a-tubulin sequences. Note that the sites at the 3' end of the a-tubulin sequences are downstream of the stop codon.

关于我们 About Us

公司简介
项目起源

基因搜项目发展历程最早可追溯至2010年01月05日域名注册，项目发起即为促进基因资源共享，避免在基因研究工作中“重复造轮子”，节约广大科研精英的宝贵时间和精力。

团队使命

让基因研究更简单!

团队愿景

您值得信赖的一站式基因研究合作伙伴!

联系我们