Expresso® Rhamnose Cloning and Expression Syst.,N-His/Expresso® Rhamnose克隆及蛋白表达系统
货号：49011-1,49011-2
规格：5 rxns,10 rxns
价格：2901,5320
产品类型：PCR 及DNA聚合酶
品牌：Lucigen
| The Expresso Rhamnose Cloning and Protein Expression Systems are designed for fast, easy, and efficient directional cloning and expression of PCR-amplified genes using Expressioneering Technology. Expressioneering Technology usesin vivohomologous recombination to seamlessly clone PCR amplified DNA into specially designed expression vectors without the need for enzymes or purification steps. A single host strain is used for both stable cloning and controlled protein expression, making Expresso Rhamnose the fastest cloning and expression systems available. The systems come complete with pre-processed expression plasmids and competent cells, supplied in single transformation vials. |
| 优点： |
| ▪Five-second, directional, enzyme-free PCR cloning! 90% recombinants!▪Single competent cell host strain for both cloning and expression.▪Tighter expression control with tunable Rhamnose (rhaPBAD) promoter. |
| 数据： |
| Figure 1. Expressioneering Technologyusesin vivohomologous recombination to seamlessly clone PCR amplified DNA into specially designed expression vectors without the need for enzymes or purification steps. The desired insert is simply amplified with primers that include 18 bases that overlap with the ends of the Expresso® vector. The unpurified PCR amplicon is then mixed with the pRham expression plasmid and the high-efficiency competent cells provided, and directly plated on appropriate media.Figure 2.pRham expression vectors.RBS, ribosome binding site; ATG, translation start site; Stop, translation end site; Kan, kanamycin resistance gene; ROP, Repressor of Priming (for low copy number); Ori, origin of replication. CloneSmart® transcription terminators (T) prevent transcription into or out of the insert, and a terminator follows the cloning site. The 6xHis affinity tag is fused to the amino terminus (pRham N-His) or at the carboxyl terminus (pRham C-His) of the expressed target protein. Also available: pRham N-His SUMO Vector for enhanced soluble protein expression with cleavable SUMO solubility tag.Figure 3. Tuning recombinant protein expression levels with rhamnose induction.The pRham C-His Kan Vector containing a gene encoding a blue fluorescent protein (BFP) was transformed intoE. cloni10G cells. An uninduced starter culture was inoculated to a starting OD600 of 0.8 into culture tubes containing LB media with 30 µg/ml kanamycin and the indicated concentrations of rhamnose (0 to 0.2% w/v) or 2% glucose. After overnight incubation at 37°C, samples were harvested by centrifugation, lysed in SDS-PAGE loading buffer, and analyzed by SDS-PAGE. The Coomassie-blue stained gel shows total cellular protein. Protein expression levels are responsive to rhamnose concentrations between 0.001% and 0.2%. |
| 相关产品： |
| ▪Expresso® Rhamnose Cloning and Expression System, C-His（货号#49012）▪ Expresso® Rhamnose SUMO Cloning and Expression System（货号#49013）▪Glucose Solution, 15% w/v（货号#49022）▪Rhamnose Solution, 20% w/v（货号#49021）▪SUMO Express Protease（货号#30801） |
技术参数 产品优点 - Five-second, directional, enzyme-free PCR cloning! 90% recombinants! - Single competent cell host strain for both cloning and expression. - Tighter expression control with tunable Rhamnose (rhaPBAD) promoter. - High throughput format friendly with autoinduction reagents.
产品应用 - The Expresso Rhamnose Cloning and Protein Expression Systems are designed for fast, easy, and efficient directional cloning and expression of PCR-amplified genes using Expressioneering Technology.
组成成分 The Expresso Rhamnose Cloning & Expression System contains pre-processed pRham N-His or pRham C-His Vector DNA, single-transformation E. cloni 10G Chemically Competent Cells (SOLOs), and the auto-induction reagents 20% Rhamnose solution and 15% Glucose solution. Also included are N-His or C-His Positive Control Insert DNAs, transformation positive control pUC DNA, and forward and reverse PCR primers to confirm clones.
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