pBAD/gIII 试剂盒

引用和文献 (4)

Invitrogen™

pBAD/gIII 试剂盒

The pBAD/gIII Kit provides all of the necessary reagents to express recombinant proteins under the tight regulation of the araBAD了解更多信息

货号	数量
V45001	1 kit

货号 V45001

价格（CNY)

15,816.00

1 kit

添加至购物车

1 kit

价格（CNY)

15,816.00

1 kit

添加至购物车

The pBAD/gIII Kit provides all of the necessary reagents to express recombinant proteins under the tight regulation of the araBAD promoter. To simplify purification, the vector pBAD/gIII encodes a leader peptide which directs the recombinant protein into the
periplasmic space. The vector provides:

• The araBAD promoter for tightly regulated expression
• Translation initiation signals for optimized E. coliexpression
• Leader peptide from the bacteriophage fd gene III protein (gIII) for secreted expression
• C-terminal polyhistidine (6xHis) tag for purification with nickel-chelating resin
• C-terminal c-myc epitope for detection and analysis with an Anti-myc Antibody

A set of three vectors is provided (A, B, and C). Each has the C-terminal tag in a different reading frame relative to the multiple cloning site to simplify in-frame cloning of your gene.

仅供科研使用。不可用于诊断程序。

耐抗生素细菌氨苄青霉素 (AmpR)

构成或诱导系统诱导

诱导剂阿拉伯糖

产品类型细菌表达载体

数量1 kit

选择试剂（真核生物）无

载体pBAD

克隆方法限制性内切酶/MCS

促进剂araBAD

蛋白标记c-Myc 抗原决定簇标签

Unit Size1 kit

内容与储存

pBAD/gIII 试剂盒包含各 20 µg pBAD/gIII A、B & C、20 µg pBAD/gIII/钙调蛋白、1 mL 20% L-阿拉伯糖、LMG194 和 TOP10 大肠杆菌穿刺培养物。在 2 - 8°C 下储存穿刺培养物。将所有其他组分储存在 -20°C 下。妥善储存时，可保证所有组分稳定储存 6 个月。

常见问题解答 (FAQ)

应使用多少L-阿拉伯糖诱导表达？

L-阿拉伯糖的用量取决于您的表达实验，我们建议在初试表达试验中采用多种不同用量的L-阿拉伯糖，范围为0.00002%至0.2%。

我应使用你们提供的TOP10细胞还是LMG194大肠杆菌菌株用于pBAD系统的表达呢？

TOP10
优点：
- 省时，可直接从克隆到表达。
- 甘油储液更稳定，因为这些菌株的基因型是endA-和recA-。
缺点：
-该菌株不是蛋白酶缺陷型的；因此，蛋白质可能被降解。

LMG194
优点：
- 可在最低限培养基中生长良好，M9除外。
- 为了表达质粒必须转化到细胞中。
-含葡萄糖的RM培养基，保证了蛋白质的低基础表达水平。
缺点：
- 不是蛋白酶缺陷型的；因此，蛋白质可能被降解。
-甘油储液可能不稳定，因为这些菌株的基因型不是recA-或endA-。

你们建议选择哪种感受态细胞株用于pBAD表达系统的表达？

我们建议使用基因型为araBADC-和araEFGH+的感受态细胞株，这种细胞可转运L-阿拉伯糖，而不会将其代谢。这对于表达研究很重要，因为在细胞内的L-阿拉伯糖水平是恒定不变的，不会随时间降低。我们可提供我们的TOP10感受态细胞或LMG194大肠杆菌菌株。

你们能否提供可用于pBAD载体中对目的基因进行测序的引物序列？

pBAD载体含有正向和反向pBAD引物，位于目的基因上下游。这些序列如下：

pBAD正向引物：5’-ATGCCATAGCATTTTTATCC-3’
pBAD反向引物：5’-GATTTAATCTGTATCAGG-3’

你们是否有pBAD载体管盖颜色列表？

pBAD/His A 红色
pBAD/His B 橙色
pBAD/His C 黄色
pBAD/His LacZ 绿色
pBad/gIII A 黄色
pBad/gIII B 绿色
pBad/gIII C 蓝色
pBAD/gIII/calmodulin 紫色

View all pBAD/gIII 试剂盒 FAQs

引用和文献 (4)

引用和文献

Abstract

Inhibition of adipocyte differentiation by resistin-like molecule alpha. Biochemical characterization of its oligomeric nature.

Authors: Blagoev Blagoy; Kratchmarova Irina; Nielsen Mogens M; Fernandez Minerva M; Voldby Jesper; Andersen Jens S; Kristiansen Karsten; Pandey Akhilesh; Mann Matthias;

Journal:J Biol Chem

PubMed ID:12189153

A novel family of cysteine-rich secreted proteins with unique tissue distribution has recently been identified. One of the members, resistin (for

The COOH Terminus of Arylamine N-Acetyltransferase from Salmonella typhimurium Controls Enzymic Activity.

Authors: Mushtaq Adeel; Payton Mark; Sim Edith;

Journal:J Biol Chem

PubMed ID:11799105

Arylamine N-acetyltransferases (NATs) are a homologous family of enzymes, which acetylate arylamines, arylhydroxylamines, and arylhydrazines by acetyl transfer from acetyl-coenzyme A (Ac-CoA) and are found in many organisms. NAT was first identified as the enzyme responsible for the inactivation of the anti-tubercular drug isoniazid in humans. The three-dimensional structure of ... More

Specificity determinants and diversification of the Brassica self-incompatibility pollen ligand.

Authors:Chookajorn T, Kachroo A, Ripoll DR, Clark AG, Nasrallah JB,

Journal:Proc Natl Acad Sci U S A

PubMed ID:14694189

Self-incompatibility in crucifers is effected by allele-specific interactions between the highly polymorphic stigmatic S locus receptor kinase (SRK) and its pollen ligand, the S locus cysteine-rich protein (SCR). Here we show that specificity in SCR function is determined by four contiguous amino acids in one variant, indicating that the minimum ... More

A general method for greatly improving the affinity of antibodies by using combinatorial libraries.

Authors:Rajpal A, Beyaz N, Haber L, Cappuccilli G, Yee H, Bhatt RR, Takeuchi T, Lerner RA, Crea R,

Journal:Proc Natl Acad Sci U S A

PubMed ID:15939870

Look-through mutagenesis (LTM) is a multidimensional mutagenesis method that simultaneously assesses and optimizes combinatorial mutations of selected amino acids. The process focuses on a precise distribution within one or more complementarity determining region (CDR) domains and explores the synergistic contribution of amino acid side-chain chemistry. LTM was applied to an ... More