pBAD/Myc-His Kit

Citas y referencias (6)

Invitrogen™

pBAD/Myc-His Kit

El kit pBAD/Myc-His proporciona todos los reactivos necesarios para extraer la proteína con una tendencia completamente regulada. El vector pBAD/Myc-HisMás información

Número de catálogo	Cantidad
V44001	1 kit

Número de catálogo V44001

Precio (CLP)

1.385.529

1 kit

Añadir al carro de la compra

Cantidad:

1 kit

Precio (CLP)

1.385.529

1 kit

Añadir al carro de la compra

El kit pBAD/Myc-His proporciona todos los reactivos necesarios para extraer la proteína con una tendencia completamente regulada. El vector pBAD/Myc-His expresa proteínas nativas o de fusión con una etiqueta C-terminal. El vector proporciona:

• El promotor de araBAD para una expresión completamente regulada
• Señales de iniciación de traducción optimizadas para la expresión de E. col
• Etiqueta de polihistidina (6xHis) C-terminal para una rápida purificación con resina quelante de níquel y detección con anticuerpos anti-His (C-terminal)
• Epítopo de c-myc C-terminal para detección y análisis con un anticuerpo anti-myc

Se proporcionan tres vectores (A, B y C). Cada uno de estos tiene la etiqueta C-terminal en un marco de lectura diferente en relación con el sitio de clonación múltiple para simplificar la clonación de genes dentro de un marco.

Para uso exclusivo en investigación. No apto para uso en procedimientos diagnósticos.

Especificaciones

Resistencia bacteriana a los antibióticosAmpicilina (AMPR)

HendiduraSitio de reconocimiento de EK (enterocinasa)

Sistema constitutivo o inducibleInducible

Agente inductorArabinosa

Tipo de productoVector de expresión bacteriana

Cantidad1 kit

Agente de selección (eucariótico)Ninguno

VectorpBAD

Método de clonaciónEnzimas de restricción/MCS

PromotoraraBAD

Etiqueta de proteínaEtiqueta de epítopo c-Myc

Unit Size1 kit

Contenido y almacenamiento

El kit pBAD/Myc-His incluye 20 μg del vector pBAD/Myc-His A, B, & C; 20 μg de pBAD/Myc-His/lacZ, medios L-arabinosa al 20%, E. coliTOP10 y LMG194. Los vectores y la L-arabinosa al 20% se deben conservar a -20 °C. Conservar los medios LMG194 y TOP10 a una temperatura de 2 a 8 °C. Se garantiza la estabilidad de todos los componentes durante 6 meses si se almacenan correctamente.

Preguntas frecuentes

How much L-arabinose should I use to induce expression with the pBAD expression system?

While the amount of L-arabinose can vary depending on your expression experiment, we suggest performing a pilot expression experiment with varying amounts of L-arabinose from 0.00002% to 0.2%.

Should I use TOP10 cells or the LMG194 E. coli strain you offer for expression with my pBAD system?

Top10

Advantages:
- Saves time, can go directly from cloning to expression.
- The glycerol stock is more stable because these strains are endA- and recA-.

Disdvantages:
- This strain is not protease-deficient. Therefore, the protein may be degraded.

LMG194

Advantages:
- Grows well in minimal media, except M9.
-Have to transform the plasmid into the cells just for expression.
-RM medium with glucose to ensure low basal level of protein.

Disadvantages:
- Not protease-deficient. Therefore, the protein may be degraded.
- The glycerol stock may not be stable because this cell strain is not recA- or endA-.

What competent cells do you recommend I use for expression with my pBAD expression system?

We recommend using a competent cell strain that is araBADC- and araEFGH+, allowing transportation of L-arabinose, but not metabolizing it. This is important for expression studies, as the level of L-arabinose will be constant inside the cell and will not decrease over time. We offer our TOP10 competent cells, or our LMG194 E. coli strain.

Can you tell me the sequence of primers that can be used in pBAD vectors to sequence my gene of interest?

The pBAD vectors contain a forward and reverse pBAD primer flanking the gene of interest. The sequences are as follows:

pBAD forward primer: 5'-ATGCCATAGCATTTTTATCC-3'

pBAD reverse primer: 5'-GATTTAATCTGTATCAGG-3'

Do you have a list of the cap colors for the pBAD vectors?

Here are the cap colors:

- pBAD/His A: Red

- pBAD/His B: Orange

- pBAD/His C: Yellow

- pBAD/His LacZ: Green

- pBad/gIII A: Yellow

- pBad/gIII B: Green

- pBad/gIII C: Blue

- pBAD/gIII/calmodulin: Purple

View all pBAD/Myc-His Kit FAQs

Citations & References (6)

Citations & References

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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