pBAD/Myc-His 试剂盒

引用和文献 (6)

Invitrogen™

pBAD/Myc-His 试剂盒

The pBAD/Myc-His Kit provides all of the necessary reagents to express your protein in a tightly regulated fashion. The pBAD/Myc-His了解更多信息

货号	数量
V44001	1 kit

货号 V44001

价格（CNY)

15,344.00

1 kit

添加至购物车

1 kit

价格（CNY)

15,344.00

1 kit

添加至购物车

The pBAD/Myc-His Kit provides all of the necessary reagents to express your protein in a tightly regulated fashion. The pBAD/Myc-His vector expresses native proteins or fusion proteins with a C-terminal tag. The vector provides:

• The araBAD promoter for tightly regulated expression
• Translation initiation signals optimized for E. coliexpression
• C-terminal polyhistidine (6xHis) tag for purification with nickel-chelating resin or detection with an Anti-His(C-term) Antibody
• C-terminal c-myc epitope for detection and analysis with an Anti-myc Antibody

Three vectors are 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)

切割EK（肠激酶）识别位点

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/Myc-His 试剂盒包含 20 µg pBAD/Myc-His A、B & C 载体、20 µg pBAD/Myc-His/lacZ、20% L-阿拉伯糖、LMG194 和 TOP10 大肠杆菌穿刺培养物。载体和 20% L-阿拉伯糖应储存于 -20°C 下。在 2-8°C 下储存 LMG194 和 TOP10 穿刺培养物。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/Myc-His 试剂盒 FAQs

引用和文献 (6)

引用和文献

Abstract

The intercellular signaling activity of the Mycobacterium tuberculosis chaperonin 60.1 protein resides in the equatorial domain.

Authors:Tormay P, Coates AR, Henderson B,

Journal:J Biol Chem

PubMed ID:15677470

'The major heat shock protein, chaperonin 60, has been established to have intercellular signaling activity in addition to its established protein-folding function. Mycobacterium tuberculosis is one of a small proportion of bacteria to encode two chaperonin 60 proteins. We have demonstrated that chaperonin 60.1 from this bacterium is a very ... More

Novel pathway for arsenic detoxification in the legume symbiont Sinorhizobium meliloti.

Authors:Yang HC, Cheng J, Finan TM, Rosen BP, Bhattacharjee H,

Journal:J Bacteriol

PubMed ID:16199569

'We report a novel pathway for arsenic detoxification in the legume symbiont Sinorhizobium meliloti. Although a majority of ars operons consist of three genes, arsR (transcriptional regulator), arsB [As(OH)3/H+ antiporter], and arsC (arsenate reductase), the S. meliloti ars operon includes an aquaglyceroporin (aqpS) in place of arsB. The presence of ... More

CopA: An Escherichia coli Cu(I)-translocating P-type ATPase

Authors:Rensing C, Fan B, Sharma R, Mitra B, Rosen BP

Journal:Proc Natl Acad Sci U S A

PubMed ID:10639134

'The copA gene product, a putative copper-translocating P-type ATPase, has been shown to be involved in copper resistance in Escherichia coli. The copA gene was disrupted by insertion of a kanamycin gene through homologous recombination. The mutant strain was more sensitive to copper salts but not to salts of other ... More

The prodrug activator EtaA from Mycobacterium tuberculosis is a Baeyer-Villiger monooxygenase.

Authors:Fraaije MW, Kamerbeek NM, Heidekamp AJ, Fortin R, Janssen DB,

Journal:J Biol Chem

PubMed ID:14610090

'EtaA is a newly identified FAD-containing monooxygenase that is responsible for activation of several thioamide prodrugs in Mycobacterium tuberculosis. It was found that purified EtaA displays a remarkably low activity with the antitubercular prodrug ethionamide. Hinted by the presence of a Baeyer-Villiger monooxygenase sequence motif in the EtaA sequence, we ... More

Genetic engineering of phytochrome biosynthesis in bacteria.

Authors: Gambetta G A; Lagarias J C;

Journal:Proc Natl Acad Sci U S A

PubMed ID:11553807

The bilin prosthetic groups of the phytochrome photoreceptors and the light-harvesting phycobiliprotein antennae arise from the oxygen-dependent ring opening of heme. Two ferredoxin-dependent enzymes contribute to this conversion: a heme oxygenase and a bilin reductase with discrete double-bond specificity. Using a dual plasmid system, one expressing a truncated cyanobacterial apophytochrome ... More

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