pBAD/His Kit
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Invitrogen™

pBAD/His Kit

El kit pBAD/His proporciona todos los reactivos necesarios para extraer la proteína con una tendencia completamente regulada. El vector pBAD/HisMás información
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Número de catálogoCantidad
V430011 kit
Número de catálogo V43001
Precio (MXN)
-
Cantidad:
1 kit
El kit pBAD/His proporciona todos los reactivos necesarios para extraer la proteína con una tendencia completamente regulada. El vector pBAD/His le permite expresar su proteína con una etiqueta N-terminal. El vector proporciona:

• El promotor de araBAD para una expresión completamente regulada
• Señales de iniciación de traducción optimizadas para la expresión de E. coli
•Etiqueta de polihistidina N-terminal (6xHis) para la purificación con resina quelante del níquel y para la detección con un anticuerpo anti-HisG
•™ Epítopo N-terminal Xpress para detección y análisis con un anticuerpo anti-Xpress™
• Sitio de escisión de enterocinasa para eliminar la etiqueta N-terminal después de la purificación

Se proporcionan tres vectores (A, B y C). Cada uno de estos tiene la etiqueta N-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 His (6x)
Unit Size1 kit
Contenido y almacenamiento
El kit pBAD/His incluye 20 μg de cada vector pBAD/His A, B & C; 20 μg de pBAD/His/lacZ, 1 ml de medios L-arabinosa al 20% y E. coli TOP10 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

Citations & References (10)

Citations & References
Abstract
Interaction between FtsZ and FtsW of Mycobacterium tuberculosis.
Authors: Datta Pratik; Dasgupta Arunava; Bhakta Sanjib; Basu Joyoti;
Journal:J Biol Chem
PubMed ID:12101218
'The recruitment of FtsZ to the septum and its subsequent interaction with other cell division proteins in a spatially and temporally controlled manner are the keys to bacterial cell division. In the present study, we have tested the hypothesis that FtsZ and FtsW of Mycobacterium tuberculosis could be binding partners. ... More
Tight Binding Inhibition of Protein Phosphatase-1 by Phosphatidic Acid. SPECIFICITY OF INHIBITION BY THE PHOSPHOLIPID.
Authors: Jones Jeffrey A; Hannun Yusuf A;
Journal:J Biol Chem
PubMed ID:11856740
'Phosphatidic acid (PA) has been identified as a bioactive lipid second messenger, yet despite extensive investigation, no cellular target has emerged as a mediator of its described biological effects. In this study, we identify the gamma isoform of the human protein phosphatase-1 catalytic subunit (PP1cgamma) as a high affinity in ... More
Biochemical properties of the PsbS subunit of photosystem II either purified from chloroplast or recombinant.
Authors: Dominici Paola; Caffarri Stefano; Armenante Franca; Ceoldo Stefania; Crimi Massimo; Bassi Roberto;
Journal:J Biol Chem
PubMed ID:11934892
'The biochemical properties of PsbS protein, a nuclear-encoded Photosystem II subunit involved in the high energy quenching of chlorophyll fluorescence, have been studied using preparations purified from chloroplasts or obtained by overexpression in bacteria. Despite the homology with chlorophyll a/b/xanthophyll-binding proteins of the Lhc family, native PsbS protein does not ... More
Genetic and biochemical properties of streptococcal NAD-glycohydrolase inhibitor.
Authors:Kimoto H, Fujii Y, Hirano S, Yokota Y, Taketo A,
Journal:J Biol Chem
PubMed ID:16380378
'The gene encoding streptolysin O (slo), a cytolysin of hemolytic streptococci, is transcribed polycistronically from the promoter of the preceding NAD-glycohydrolase (NADase) gene (nga). Between nga and slo, a putative open reading frame (orf1) is located whose function has been totally unknown. Present investigation demonstrated that the orf1 encodes a ... More
Assimilation of nicotinamide mononucleotide requires periplasmic AphA phosphatase in Salmonella enterica.
Authors:Grose JH, Bergthorsson U, Xu Y, Sterneckert J, Khodaverdian B, Roth JR,
Journal:J Bacteriol
PubMed ID:15968063
'Salmonella enterica can obtain pyridine from exogenous nicotinamide mononucleotide (NMN) by three routes. In route 1, nicotinamide is removed from NMN in the periplasm and enters the cell as the free base. In route 2, described here, phosphate is removed from NMN in the periplasm by acid phosphatase (AphA), and ... More