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仪器信息网试剂标物生化试剂生物分子 pEGFP-1哺乳启动子检测质粒

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pEGFP-1哺乳启动子检测质粒

价格：￥ 980

品牌：钦诚生物

供货周期：现货

货号：QCP0132

规格：2UG

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详细信息

pEGFP-1哺乳启动子检测质粒

基本信息

复制子: F1 ori , pUC ori

终止子: SV40 poly(A) signal

质粒分类: 哺乳系列质粒；哺乳荧光质粒；哺乳绿色质粒

质粒大小: 4151bp

质粒标签: C-EGFP

原核抗性: 卡那霉素Kan（50μg/ml）

筛选标记: 新霉素Neo/G418

克隆菌株: DH5α等大肠杆菌

培养条件: 37℃，有氧 LB

表达宿主: 293T等哺乳细胞

培养条件: 37℃，5%CO2

诱导方式: 无须诱导，瞬时表达

5'测序引物: EGFP-N（CGTCGCCGTCCAGCTCGACCAG）

3'测序引物: EGFP-F (CCAGCAACGCGGCCTTTTTA)

质粒简介

pEGFP-1编码野生型GFP（1-3）的红移型变体，其已经针对更明亮的荧光和哺乳动物细胞中更高的表达进行了优化。（激发最大值= 488nm;发射最大值= 507nm）pEGFP-1骨架还提供了用于在大肠杆菌中繁殖的pUC起始点和用于单链DNA生产的f1起源。

pEGFP-1 encodes a red-shifted variant of wild-type GFP (1–3) which has been optimized for brighter fluorescence and higher expression in mammalian cells. (Excitation maximum = 488 nm; emission maximum = 507 nm.) pEGFP-1 encodes the GFPmut1 variant (4) which contains the double-aminoacid substitution of Phe-64 to Leu and Ser-65 to Thr. The coding sequence of the EGFP gene contains more than 190 silent base changes which correspond to human codon-usage preferences (5). Sequences flanking EGFP have been converted to a Kozak consensus translation initiation site (6) to further increase the translation efficiency in eukaryotic cells. pEGFP-1 is a promoterless EGFP vector which can be used to monitor transcription from different promoters and promoter/enhancer combinations inserted into the MCS located upstream of the EGFP coding sequence. SV40 polyadenylation signals downstream of the EGFP gene direct proper processing of the 3' end of the EGFP mRNA. The vector backbone also contains an SV40 origin for replication in mammalian cells expressing the SV40 T antigen. A neomycin-resistance cassette (Neor) allows stably transfected eukaryotic cells to be selected using G418. The Neor cassette consists 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. A bacterial promoter upstream of this cassette confers kanamycin resistance in E. coli. The pEGFP-1 backbone also provides a pUC origin of replication for propagation in E. coli and an f1 origin for single-stranded DNA production.

EGFP can be used as an in vivo reporter of gene expression . Promoters should be cloned into the pEGFP-1 MCS upstream from the EGFP coding sequences. Without the addition of a functional promoter, this vector will not express EGFP. The recombinant EGFP vector can be transfected into mammalian cells using any standard transfection method. If required, stable transformants can be selected using G418 .

质粒图谱

pEGFP-1哺乳启动子检测质粒使用说明：

1、收到质粒干粉后请先5000rpm离心1min，再加入20μl无菌水溶解质粒，室温放置1min；

2、从-80℃冰箱中取出相应的感受态，置于冰盒上解冻，并做好标记；

3、取2μl质粒加至100μl感受态中，冰浴30min；

4、42℃热激90s，再冰浴2min；

5、加入900μl无抗的LB液体培养基，180rpm震荡培养45min；

6、6000rpm离心5min，仅留100ul上清混匀菌体沉淀；

7、混匀后的菌液加至对应抗性的LB平板上，倒入适量玻璃珠，涂匀液体；

8、将平板正向培养1h，再倒置培养12h~16h；

9、挑取单克隆菌落至对应抗性的LB液体培养基中，震荡培养12h~16h，根据实验需要提取质粒。

pEGFP-1哺乳启动子检测质粒注意事项：

1、如果您收到的是甘油菌种，请先四区划线，挑取单克隆培养。

2、如果第二天转化平板长的过多，请将质粒按比例稀释后再转化。

生化试剂 pEGFP-1哺乳启动子检测质粒由上海钦诚生物科技有限公司为您提供，如想了解更多关于生物分子 pEGFP-1哺乳启动子检测质粒的报价、规格、厂家等信息，欢迎来电或留言咨询。除供应pEGFP-1哺乳启动子检测质粒外，上海钦诚生物科技有限公司还可为您提供： HRMVPCS 永生化人视网膜周细胞 QCH791、 L363 人多发性骨髓瘤细胞 QCH792、 Colo680N 人食管鳞癌细胞 QCH793

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