E. coli S30 Extract System for Circular DNA

产品编号：L1020 品牌：promega 原厂货号：L1020
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℃

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其它组分：

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

S30 Extract, Circular【子货号：L464A，包装：3 x 150μl，，运保温度：-70℃】

Luciferase Assay Reagent【子货号：L486A，包装：1 x 250μl，，运保温度：-70℃】

Luciferase Dilution Reagent【子货号：L487A，包装：1 x 1ml，，运保温度：-70℃】

pBESTluc™ DNA, Circular (Control)【子货号：L492A，包装：1 x 10μg，，运保温度：-70℃】

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

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

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

描述：

说明

E. coli S30 Extract for Circular DNA( 用于环状DNA 模板的E. coli S30 提取物系统) 简化了在质粒或λ 载体中克隆的DNA序列的转录和翻译，提供了鉴定和研究多肽特征的有力工具。研究者只需准备含有合适的原核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OmpT 内蛋白酶和lon 蛋白酶活性缺失的大肠杆菌B 株制得。从而极大地增强了表达蛋白的稳定性，因为若在体内翻译的话，表达的目标蛋白将会被蛋白酶降解。由于宿主编码的抑制物的存在，某些蛋白在体内表达的水平很低，使用S30 体外系统可提高这些蛋白的翻译水平。

特点

.  更好的稳定性：降低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( 氨基酸除外)，如核糖体，各种tRNAs，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 S30 Extract for Circular DNA simplifies the transcription/translation of DNA sequences cloned in plasmid or λ vectors, providing a powerful tool for identifying and characterizing polypeptides. The investigator needs only to supply the cloned DNA containing the appropriate prokaryotic promoter and ribosome binding sites. The S30 Extract for Circular DNA Templates 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 a greater stability of expressed proteins that would otherwise be degraded by proteases if expressed in vivo. The S30 in vitro system also allows higher expression levels of proteins that are normally expressed at low levels in vivo due to the action of host-encoded repressors.

Features - Benefits

Greater Stability: Reduced chance of expressed proteins degrading.
Complete: Contains all necessary components for coupled transcription/translation.
Low Background: System 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

Regulation of prokaryotic gene expression.
Identification of cloned DNA gene products or mapping of polypeptides to defined DNA fragments.
Rapid screening for translation from eukaryotic or prokaryotic DNAs cloned in vectors designed for high expression in E. coli.
Screening of compounds for inhibition of bacterial translation.

For more information, see the Protocols & Applications Guide.

References

Zubay, G. (1973) Ann. Rev. Genet. 7, 267–87.
Zubay, G. (1980) Meth. Enzymol. 65, 856–77.
Pratt, J.M. (1984) In: Transcription and Translation, Hames, B.D. and Higgens S.J., eds., IRL Press, Oxford, 179.
Studier, F.W. and Moffatt, B.A. (1986) J. Mol. Biol. 189, 113–30.
Collins, J. (1979) Gene 6, 29–42.