Bac-to-Bac™ Vector Kit

Citas y referencias (12)

Gibco™

Bac-to-Bac™ Vector Kit

El kit de vector Bac-to-Bac™ contiene un vector pFastBac™ 1, así como un vector de control de expresión, destinado paraMás información

Número de catálogo	Cantidad
10360014	1 kit

Número de catálogo 10360014

Precio (MXN)

Cantidad:

1 kit

El kit de vector Bac-to-Bac™ contiene un vector pFastBac™ 1, así como un vector de control de expresión, destinado para su uso como parte del sistema de expresión de baculovirus Bac-to-Bac™ (n.º de cat. 10359-016), que permite la producción eficaz de baculovirus recombinante para la prueba de expresión en células de insectos. El sistema Bac-to-Bac™ se basa en la generación de baculovirus recombinante mediante la transposición específica del sitio en E. coli en lugar de recombinación homóloga en células de insectos (Figura 1). Características del sistema:

• Bácmido de expresión que ahorra tiempo. Con Bac-to-Bac™, el casete de expresión del vector pFastBac™ se recombina con el bácmido principal en células competentes DH10Bac™ de E. coli (no incluidas en este kit vectorial) para formar un bácmido de expresión. A continuación, el bácmido se transfecta en células de insectos para producir partículas recombinantes de baculovirus.
• Fácil detección de colonias El bácmido parental en E. coli DH10Bac™ contiene un segmento del gen lacZα. El gen lacZα se interrumpe tras la transposición del casete de expresión en el bácmido permitiendo la selección azul/blanca de recombinantes. Esto facilita la identificación de colonias recombinantes.
El sistema de expresión de baculovirus Bac-to-Bac™ está diseñado para la producción rápida y a pequeña escala de baculovirus recombinantes. El vector pFastBac™ 1 proporcionado por este kit ofrece el promotor fuerte de poliedrina para la expresión de proteínas y gran sitio de clonación múltiple para clonación simplificada.

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

Especificaciones

Tipo de productoKit de vectores

Cantidad1 kit

VectorpFastBac

Método de clonaciónEnzimas de restricción/MCS

Línea de productosBac-to-Bac

PromotorPoliedrina

Etiqueta de proteínaSin etiquetar

Unit SizeEach

Contenido y almacenamiento

Contiene 10 μg de vector pFastBac™1 y vector de control pFastBac™ 1-Gus.

Conservar a -20° C.

Preguntas frecuentes

I cannot grow this white colony in liquid culture. What should I do?

The concentration of gentamicin might be too high. Try lowering the amount to 5 µg/mL and try adding more of the colony to the culture medium.

What has happened when I see blue colonies? How about colonies which are blue in the center and white on the edges?

In the case of a blue colony, the E. coli has the bacmid and the plasmid in it, allowing the cells to survive the selection process. However, because the transposition has not occurred, the LacZ gene is not disrupted. For bulls-eye colonies, this indicates that the transposition took place when the colony was growing. Re-streaking for an isolated clone from the white portion of the mixed colony should yield some colonies where transposition occurred.

I'm getting mostly white/wild-type plaques instead of blue/recombinant plaques. What am I doing wrong?

This is typically an indication of poor homologous recombination. Check the plasmid/linear DNA ratio you used. If there are some blue plaques, however, expand those viruses and check for their protein. In our experience, they are correct, even if they were in relatively low abundance.

I've infected my cells and see large polyhedra in one cell and smaller polyhedra (more numerous) in a neighboring cell. Is this normal?

Yes, cells are infected with wild-type virus individually and will develop polyhedra at different rates until all the cells in the flask are infected. The polyhedra in cells will form in approximately 3-4 days, differing in size and number until they reach their maximum capacity and burst the cell, releasing tiny particles of virus into the medium.

I'm worried that I am not getting plaques. How many days does it take to see plaques and what size are they typically?

Normally, very small white dots show up about 5-7 days and 1 mm plaques show up around day 10. Plaques can vary in size from 1 mm to 4 mm.

View all Bac-to-Bac™ Vector Kit FAQs

Citations & References (12)

Citations & References

Abstract

Cytoskeletal changes regulated by the PAK4 serine/threonine kinase are mediated by LIM kinase 1 and cofilin.

Authors: Dan C; Kelly A; Bernard O; Minden A;

Journal:J Biol Chem

PubMed ID:11413130

'PAK4 is the most recently identified member of the PAK family of serine/threonine kinases. PAK4 differs from other members of the PAK family in sequence and in many of its functions. Previously, we have shown that an important function of this kinase is to mediate the induction of filopodia in ... More

Allocation of helper T-cell epitope immunodominance according to three-dimensional structure in the human immunodeficiency virus type I envelope glycoprotein gp120.

Authors: Dai G; Steede N K; Landry S J;

Journal:J Biol Chem

PubMed ID:11551929

'The specificity and intensity of CD4(+) helper T-cell responses determine the effectiveness of immune effector functions. Promiscuously immunodominant helper T-cell epitopes in the human immunodeficiency virus (HIV) envelope glycoprotein gp120 could be important in the development of broadly protective immunity, but the underlying mechanisms of immunodominance and promiscuity remain poorly ... More

Interaction of Fibroblast Growth Factor Receptor 3 and the Adapter Protein SH2-B. A ROLE IN STAT5 ACTIVATION.

Authors: Kong Monica; Wang Ching S; Donoghue Daniel J;

Journal:J Biol Chem

PubMed ID:11827956

'Fibroblast growth factor receptor 3 (FGFR3) influences a diverse array of biological processes, including cell growth, differentiation, and migration. Activating mutations in FGFR3 are associated with multiple myeloma, cervical carcinoma, and bladder cancer. To identify proteins that interact with FGFR3 and which may mediate FGFR3-dependent signaling, a yeast two-hybrid screen ... More

Functional differences between the human ATP-dependent nucleosome remodeling proteins BRG1 and SNF2H.

Authors: Aalfs J D; Narlikar G J; Kingston R E;

Journal:J Biol Chem

PubMed ID:11435432

'ATP-dependent nucleosome remodeling complexes can be grouped into several classes that may differ in their biochemical remodeling activities and biological roles. Although there are a number of biochemical studies of each class of remodeler, there are very little data directly comparing the biochemical activities of remodelers from different classes. We ... More

PIAS1 and PIASxalpha function as SUMO-E3 ligases toward androgen receptor and repress androgen receptor-dependent transcription.

Authors:Nishida T, Yasuda H,

Journal:J Biol Chem

PubMed ID:12177000

The androgen receptor (AR) has been shown to be modified by SUMO-1, a ubiquitin-like protein. Recently we showed that PIAS family proteins function as SUMO-E3 ligases. Here we provide evidence that PIAS1 and PIASxalpha act as specific SUMO-E3 ligases for the AR. PIAS1 and PIASxalpha but not PIAS3 or PIASxbeta ... More

View all Bac-to-Bac™ Vector Kit citations