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lucigen 49003-1 Expresso? T7 SUMO Cloning
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產品簡介：北京中北林格供應lucigen 49003-1 Expresso? T7 SUMO Cloning，中北林格是epicentre總代理，提供epicentre產品貨號報價查詢。其他包括lucigen總代理，cellscript總代理，Biosearch technologies總代理，epicentre總代理，illumina總代理，ICLLAB總代理，ImmunoReagents總代理。
產品名稱:Expresso? T7 SUMO Cloning and Expression System
貨號:49003-1
規格:5 rxns
存儲溫度:-20o & -80o C
品牌：lucigen / epicentre
供應商：中北林格
產地：us
發貨地：北京
lucigen 49003-1 Expresso? T7 SUMO Cloning 產品說明書
產品描述
可以快速，容易的克隆和表達困難蛋白。是基于定向PCR克隆系統所研制的克隆表達系統。
Expresso saves you a day!
完整的克隆表達系統
Expresso? T7 克隆和蛋白表達系統是研制出的一款快速，簡單，有效的定向克隆和表達PCR擴增產物的系統。系統含有預處理的pETite? N或C端組氨酸標簽克隆載體，兩個感受態細胞，存在于一個單獨的轉化瓶中。高效HI-Control? 10G化學感受態細胞能穩定克隆，HI-Control BL21(DE3)感受態細胞提供了嚴謹的可控蛋白表達。因此可以避免泄露型T7啟動子系統存在的表達問題。N或C端的組氨酸標簽蛋白可以使你快速的使用鎳柱純化表達蛋白。系統還具有在N端編碼6xHis SUMO融合標簽可以改善表達蛋白的可溶性。其他表達系統為鼠李糖啟動子控制的表達。
5秒定向克隆PCR擴增基因
Expresso? T7 克隆和蛋白表達系統使用Expressioneering ?技術，是一個無酶的重組克隆策略到無縫基因載體整合過程。使用帶有18bp載體互補序列的引物對靶向基因進行PCR擴增。不同于其他的限制酶方法或無連接酶方法，不需要進一步純化和酶處理PCR產物。只需要簡單混合1μl不需純化的PCR反應產物至預處理的pETite? T7表達載體，立即轉化至HI-Control? 10G化學感受態細胞（如圖1）。
優點：
省時，無酶的克隆體系，利用克隆載體和細胞只需數秒完成。
高效：?90%的重組子。
嚴謹控制表達，N端或C端帶有6組氨酸標簽蛋白。
Expressioneering? 技術，所有Expresso?產品的特征為無連接酶克隆。
克隆載體構建，使研究時間節省數十小時
全新New pETite T7 載體包含:
用于高表達的T7強啟動子
用于蛋白快速純化的位于N端或C端的組氨酸蛋白融合標簽。
大小只有2.2kb，易于下游操作。
專利克隆技術增加了克隆效率。
lucigen 49003-1 Expresso? T7 SUMO Cloning 圖例說明
pETite vector maps
Figure 2. pETite T7 expression vectors: Small size (2.2 kb vs. 5.4 kb for pET) for easier manipulation, including targeted mutagenesis. Vectors are pre-processed for instant enzyme-free cloning of PCR products with a choice of amino-terminal or carboxyl-terminal fusion of 6xHis tag to protein of interest.
使用HI-Control 10G 化學感受態細胞獲得高效率克隆
高效的HI-Control 10G 化學感受態細胞可以確保篩選得到插入正確的克隆。對于大多數基因，> 90%的克隆都是靶向基因插入方向正確的陽性克隆（如圖三）。
High cloning efficiency with Expresso T7 System
Figure 3. High cloning efficiency with Expresso T7 System.
Pre-processed pETite C-His vector was mixed with 1 μl of unpurified PCR gene product and transformed into HI-Control 10G Chemically Competent Cells. Colony PCR was performed on 18 randomly chosen colonies; 17 of 18 contained insert of the correct size.
HI-Control BL21(DE3)細胞控制泄漏蛋白表達
HI-Control BL21(DE3) 細胞含有高水平lac抑制物，可以保證嚴謹控制T7
RNA聚合酶的表達。嚴格控制意味著更好的適應潛在毒性基因產物（圖4）。
Expresso vs. pET28a
Figure 4. Expression of various proteins using the Expresso T7 System versus a pET Vector. Genes encoding a DNA polymerase, a blue fluorescent protein (BFP), or ATP synthase b subunit (membrane protein) were cloned into pET28a or pETite vectors with N-terminal or C-terminal 6xHis tags (as indicated). pET28a clones were transformed into standard BL21(DE3) cells, and pETite clones were transformed into HI-Control BL21(DE3) cells for expression. Cultures were grown in LB at 37°C to an OD600 of 0.5 to 0.7 (odd-numbered lanes) and induced for 3 hours with 1 mM IPTG (even-numbered lanes). Cells were pelleted and lysed directly in SDS-PAGE loading buffer, and 0.05 OD equivalents were analyzed by gel electrophoresis. The gel was stained with Coomassie blue.
Expression and purification of active soluble fluorescent protein.
Figure 5. Purification of a 6xHis tagged fluorescent protein. HI-Control BL21(DE3) cells harboring pETite C-His vector containing a yellow fluorescent protein (YFP) gene were grown at 37°C in LB media to an OD600 of 0.6 (lane 1), then induced with
1 mM IPTG for 4 hours (lane 2). Cells were harvested and lysed by sonication in 300 mM NaCl, 50 mM Tris-HCl pH 8.0. Cleared lysate was loaded onto an Ni-NTA Sepharose? Column. Column flow-through (lane 3, FT) and wash (lane 4, W) fractions were collected. The bound YFP, shown in the column at left, was eluted with wash buffer containing 300 mM imidazole (lanes 5-12, E1-E8). Fluorescent fractions align with the pure protein as shown on the gel.
lucigen 49003-1 Expresso? T7 SUMO Cloning 貨號詳情
Contact your local distributor for pricing
Product Description Size Cat. No.
Expresso? T7 Cloning and Expression System, N-His 5 rxns 49001-1
10 rxns 49001-2
Expresso? T7 Cloning and Expression System, C-His 5 rxns 49002-1
10 rxns 49002-2
Expresso? T7 Cloning and Expression System, N/C-His Combo, 5 of each of N-His and C-His 10 rxns 49000-1
Expresso? T7 SUMO Cloning and Expression System 5 rxns 49003-1
10 rxns 49003-2
HI-Control? 10G Chemically Competent Cells (SOLOs) 12 rxns (SOLOs) 60110-1
HI-Control? BL21(DE3) Chemically Competent Cells (SOLOs) 12 rxns (SOLOs) 60435-1
SUMO Express Protease 200 U 30801-2
ORDER INFORMATION
T7克隆和表達系統含有預處理的pETite? N-His and/or pETite C-His 載體DNA, 用于克隆的HI-Control? 10G化學感受態細胞和用于蛋白表達的HI-Control BL21(DE3) 化學感受態細胞。同時還包括N端或C端組氨酸標簽陽性對照插入DNA和轉化陽性對照PUC DNA .
T7 SUMO克隆和表達系統含有預處理的
pETite? N-His SUMO 載體 DNA, 用于克隆的HI-Control? 10G 化學感受態細胞和用于蛋白表達的 HI Control BL21(DE3) 化學感受態細胞，也包括SUMO 陽性對照插入DNA, 轉化陽性對照 pUC DNA, SUMO表達蛋白酶, SUMO 切割控制蛋白, 鑒定克隆的正反向PCR引物。
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北京中北林格科技發展有限公司
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- 北京市昌平區北清路生命科學園北清創意園2-1-102
