pBAD/gIII Kit
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Citas y referencias (4)
Invitrogen™

pBAD/gIII Kit

El kit pBAD/gIII proporciona todos los reactivos necesarios para la expresión de proteínas recombinantes bajo una regulación completa del promotorMás información
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Número de catálogoCantidad
V450011 kit
Número de catálogo V45001
Precio (MXN)
-
Cantidad:
1 kit
El kit pBAD/gIII proporciona todos los reactivos necesarios para la expresión de proteínas recombinantes bajo una regulación completa del promotor de araBAD. Para simplificar la purificación, el vector pBAD/gIII codifica un péptido líder que dirige la proteína recombinante al
espacio periplásmico. El vector proporciona:

• El promotor de araBAD para una expresión completamente regulada
• Señales de iniciación de traducción para una expresión de E. col optimizada
• Péptido líder de la proteína del gen fd III bacteriófaga (gIII) para la expresión secretada
• Etiqueta de polihistidina (6xHis) C-terminal para purificación con resina quelante de níquel
• Epítopo de c-myc C-terminal para detección y análisis con un anticuerpo anti-myc

Se proporciona un conjunto de 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)
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/gIII incluye 20 μg de cada vector pBAD/gIII A, B, & C; 20 μg de pBAD/gIII/calmodulin, 1 ml de medios L-arabinosa al 20%, E. coli TOP10 y LMG194. Conservar los medios a una temperatura entre 2 y 8 °C. Conservar todos los demás componentes a -20 °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/gIII Kit FAQs

Citations & References (4)

Citations & References
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