E. coli T7 S30 Extract System for Circular DNA

产品编号：L1130 品牌：promega 原厂货号：L1130
产品分类：蛋白质表达与纯化 > 蛋白质表达系统 > 无细胞表达系统
应用分类：

包装：

30 reactions

运保温度:

–70°C

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其它组分：

T7/S30 Extract for Circular DNA【子货号：L114A，包装：3 x 150μl，，运保温度：–70°C】

Amino Acid Mixture Minus Cysteine【子货号：L447A，包装：1 x 175μl，，运保温度：–70°C】

S30 Premix without Amino Acids【子货号：L512A，包装：1 x 750μl，，运保温度：–70°C】

Amino Acid Mixture Minus Leucine【子货号：L995B，包装：1 x 175μl，，运保温度：–70°C】

Amino Acid Mixture Minus Methionine【子货号：L996B，包装：1 x 175μl，，运保温度：–70°C】

PinPoint™ Xa Control Vector【子货号：V204A，包装：1 x 5μg，，运保温度：–70°C】

描述：

说明

E. coli T7 S30 Extract System for Circular DNA( 用于环状DNA 模板的E. coli T7 S30 提取物系统) 通过提供含有用于转录的RNA 聚合酶和所有翻译所必需组分的提取物，简化了克隆到质粒或含有T7 启动子的λ 载体中的DNA 序列的转录和翻译。研究者只需提供含有T7 启动子和核糖体结合位点的克隆DNA 即可。这种提取物的制备采用并改进了Zubay 描述的方法，是从Omp T 内蛋白酶和lon 蛋白酶活性缺失的大肠杆菌B 株制得。这极大地增强了翻译蛋白的稳定性，否则，若在体内翻译的话，产物蛋白将会被蛋白酶降解。

特点

. 灵活：能使用含有T7 启动子和一个核糖体结合位点的任何克隆进行翻译，而其它的S30 提取物需要大肠杆菌启动子才可进行翻译。

• 稳定性强：减少了表达蛋白被降解的机会。

• 完备：包含了偶联的转录/ 翻译反应需要的所有组分。

• 低背景：系统合成的内源蛋白质水平极低。

• 经过优化：每批S30 提取物的预混合物已经经过优化，包括所有其它必需组分( 除氨基酸外)，如核糖体，各种tRNA，PEP( 磷酸烯醇丙酮酸) 和盐类。

• 可定制或批量订购：请访问www.promega.com/myway/，了解该产品的更多定制信息。

原厂资料：

The E. coli T7 S30 Extract System for Circular DNA simplifies the transcription/translation of DNA sequences cloned in plasmid or λ vectors containing a T7 promoter by providing an extract that contains T7 RNA polymerase for transcription and all components needed for translation. The investigator only supplies cloned DNA containing a T7 promoter and a ribosome binding site. This product is prepared by modifications of the method described by Zubay from an E. coli strain B deficient in OmpT endoproteinase and lon protease activity. This results in greater stability of expressed proteins that would otherwise be degraded by proteases if expressed in vivo.

Features - Benefits

Flexible: Can translate using any clone that has a T7 promoter and a ribosome binding site. Other S30 extracts require an E. coli promoter.
Greater Stability: Reduced chance of expressed proteins degrading.
Complete: Contains all components needed for coupled transcription/translation.
Low Background: Synthesizes very low levels of endogenous proteins.
Optimized: Premix is optimized for each lot of S30 Extract and contains all other required components (except amino acids), such as ribonucleotides, tRNAs, PEP (phosphoenol pyruvate) and salts.
Choose Your Configuration: Learn more about our custom options for this product at: www.promega.com/custom/

Applications

Synthesis of radiolabeled protein.
Verification of cloned gene expression before in vivo protein expression in E. coli.
Use of protein in functional studies of transcription and translation.
Use of protein as a tracer in protein purification.
Incorporation of unnatural amino acids into proteins for structural studies.

For more information, see the Protocols & Applications Guide.

References

Nevin, D.E. and Pratt, J.M. (1991) FEBS Lett. 291, 259–63.
Zubay, G. (1973) Ann. Rev. Genet. 7, 267–87.
Zubay, G. (1980) Meth. Enzymol. 65, 856–77.
Studier, F.W. and Moffatt, B.A. (1986) J. Mol. Biol. 189, 113–30.
Pratt, J.M. (1984) In: Transcription and Translation, Hames, B.D. and Higgens S.J., eds., IRL Press, Oxford, 179.