Shop All Insect & Baculovirus Expression Vectors

pIB/V5-His-DEST Vector (Invitrogen™)

The pIB/V5-His-DEST vector is designed for rapid lambda phage site-specific recombination with a Gateway® entry clone and subsequent expression in Sf9, Sf21, or High Five™ insect cells. As part of the InsectSelect™ System, pIB/V5-His-DEST features:

• The OpIE2 promoter for constitutive expression
• Blasticidin resistance gene driven by the weak GP64 promoter for selection of stable cell lines that have integrated the plasmid into a transcriptionally active site
• C-terminal V5 epitope and polyhistidine (6xHis) sequence for detection with Invitrogen's Anti-V5 Antibody and easy purification with a nickel-chelating resin

pAc5.1/V5-His A, B, & C Vectors (Invitrogen™)

pAC5.1/V5-His vectors are designed for use in Drosophila cells to achieve high-level transient expression of recombinant proteins. The vector offers the strong, constitutive promoter from the Drosophila actin 5C gene. The pAC5.1/V5-His vectors can be used with the DES™-Inducible Kits (K512001 and K412001) for constitutive expression of your protein of interest. The pAc5.1/V5-His vectors also offer the following features:

• Small size (5.4 kb) to improve DNA yields and increase subcloning efficiency
• C-terminal V5 epitope tag for rapid detection with Invitrogens Anti-V5 Antibody
• C-terminal 6xHis tag for simple purification of recombinant fusion proteins using nickel-chelating resin

To facilitate cloning, a set of three vectors-A, B, and C-is provided. Each vector has the multiple cloning site in a different reading frame relative to the coding sequence of the C-terminal tag.

InsectSelect™ Glow Kit with Sf9 Cells (Invitrogen™)

The InsectSelect™ Glow Kit features the pIZT/V5-His vector for high-level expression of your
gene of interest. This vector has the following features:

• The OpIE2 promoter for constitutive expression
• The Zeocin™ resistance gene for rapid selection of stably transfected cell lines
• C-terminal V5 epitope and polyhistidine (6xHis) sequence for detection with Invitrogen's Anti-V5 Antibody and rapid purification with nickel-chelating resin

In addition, pIZT/V5-His expresses a fusion of the green fluorescent protein and the Zeocin™ resistance
protein (Zeo-GFP). The Zeo-GFP fusion protein permits rapid selection of stably transfected cell lines
with Zeocin™ and confers a fluorescent phenotype that simplifies identification of transfected cells.

DES™-Inducible Kit with pCoBlast (Invitrogen™)

The DES®-Inducible Kit provides the expression vector pMT/V5-His and a choice of Blasticidin (pCoBlast) or hygromycin (pCoHygro) selection. This vector uses the Drosophila metallothionein gene promoter that is induced in S2 cells upon addition of copper sulfate or cadmium chloride to the culture medium. The pMT/V5-His vector offers the following features:

• Small size (3.5 kb) to improve DNA yields and increase subcloning efficiency
• C-terminal V5 epitope tag for rapid detection with Invitrogen's Anti-V5 Antibody
• C-terminal 6xHis tag for simple purification of recombinant fusion proteins using nickel-chelating resin

To facilitate cloning, a set of three vectors-A, B, and C-is provided. Each vector has the multiple cloning site in a different reading frame relative to the coding sequence of the C-terminal tag.

Bac-N-Blue™ Transfection Kit (Invitrogen™)

The Bac-N-Blue™ Linear DNA was specifically designed for recombination with the pBlueBac vectors and pMelBac. Recombinant viruses have a full-length, functional lacZ gene resulting in the production of
blue plaques. This allows for easy identification and purification. Bac-N-Blue™ DNA can be used with any polyhedrin promoter-based baculovirus transfer vector. Bac-N-Blue™ DNA is linearized at three
sites, one of which is in a gene that is essential for viral propagation (1). This leads to a decrease in non-recombinant virus, making selection and purification of recombinant virus easy. In addition to the viral DNA, the Bac-N-Blue™ Transfection Kit contains Cellfectin® Reagent for high transfection efficiency.

Bac-to-Bac™ HT Vector Kit (Gibco™)

The Bac-to-Bac® HT Vector is designed for use as part of the Bac-to-Bac® Baculovirus Expression System (Cat. No. 10359-016) for the expression and purification of histidine-tagged recombinant proteins in Sf9, Sf21, or High Five™ Cells following bacmid generation in E. coli. The pFastBac™ HT vector offers the following features:

• Strong polyhedrin promoter for protein expression
• Three reading frames for simplified cloning
• N-terminal 6xHis tag for simple purification of recombinant fusion proteins
• TEV protease cleavage site for removal of the histidine tag following protein purification

Bac-to-Bac™ Baculovirus Expression System (Gibco™)

The Bac-to-Bac baculovirus expression system enables the efficient production of recombinant baculovirus for expression testing in insect cells. The system relies on generation of recombinant baculovirus by site-specific transposition in E. coli rather than homologous recombination in insect cells. This system features:

Time-saving expression bacmid—With the Bac-to-Bac system, the expression cassette of the pFastBac vector recombines with the parent bacmid in DH10Bac E. coli Competent Cells to form an expression bacmid. The bacmid is then transfected into insect cells for production of recombinant baculovirus particles.

Easy colony screening—The parent bacmid in DH10Bac E. coli contains a segment of the lacZa gene. The lacZa gene is disrupted upon transposition of the expression cassette into the bacmid, allowing for blue/white selection of recombinants for easier identification of recombinant colonies.

High transfection efficiency with ExpiFectamine Sf Transfection Reagent—The Bac-to-Bac TOPO expression kits now come with the next-generation ExpiFectamine Sf Transfection Reagent for efficient DNA transfection in insect cells using fast, flexible protocols. Find out more about this reagent ›

Flexibility—The Bac-to-Bac system comes with a pFastBac vector that contains a large multiple cloning site (MCS) for simplified cloning. Two additional vector formats are offered separately: pFastBac HT vector for production of histidine-tagged recombinant proteins and pFastBac Dual vector for expression of two proteins simultaneously using the p10 and polyhedrin promoters.

High protein expression—The pFastBac vector uses the strong polyhedrin promoter to generate high levels of expression in a variety of insect cell line such as Sf9, Sf21, and High Five cells.

pIZT/V5-His Vector Kit (Invitrogen™)

The InsectSelect™ Glow Kit features the pIZT/V5-His vector for high-level expression of your
gene of interest. This vector has the following features:

• The OpIE2 promoter for constitutive expression
• The Zeocin™ resistance gene for rapid selection of stably transfected cell lines
• C-terminal V5 epitope and polyhistidine (6xHis) sequence for detection with Invitrogen's Anti-V5 Antibody and rapid purification with nickel-chelating resin

In addition, pIZT/V5-His expresses a fusion of the green fluorescent protein and the Zeocin™ resistance
protein (Zeo-GFP). The Zeo-GFP fusion protein permits rapid selection of stably transfected cell lines
with Zeocin™ and confers a fluorescent phenotype that simplifies identification of transfected cells.

DES™-Inducible/Secreted Kit with pCoHygro (Invitrogen™)

The DES®-Inducible/Secreted Kit provides the vector pMT/BiP/V5-His for inducible, secreted expression of recombinant proteins and a choice of Blasticidin (pCoBlast) or hygromycin (pCoHygro) selection. This vector offers the inducible metallothionein promoter that is induced upon addition of copper sulfate or cadmium chloride. The N-terminal signal sequence from the insect BiP gene is provided to direct the recombinant fusion protein through the secretory pathway of S2 cells into the culture medium. The pMT/BiP/V5-His vector offers the following additional features:

• Small size (3.6 kb) to improve DNA yields and increase subcloning efficiency
• C-terminal V5 epitope tag for rapid detection with Invitrogens Anti-V5 Antibody
• C-terminal 6xHis tag for simple purification of recombinant fusion proteins using nickel-chelating resin

To facilitate cloning, a set of three vectors-A, B, and C-is provided. Each vector has the multiple cloning site in a different reading frame relative to the BiP signal sequence.

BaculoDirect™ C-Term Transfection Kit (Invitrogen™)

The BaculoDirect™ Baculovirus Expression System is a powerful and versatile eukaryotic system for high-level protein expression in insect cells. The combination of Gateway™ Technology with baculovirus expression makes the BaculoDirect System the fastest and easiest method for production of recombinant baculovirus.

How it works
BaculoDirect™ Linear DNA (the baculovirus genome) is engineered to include attR sites for quick and efficient recombination with a Gateway entry clone. The gene of interest is recombined from the entry clone into the BaculoDirect Linear DNA using a simple, one-hour LR reaction (Figure 1). The resulting reaction mix contains the recombinant baculovirus carrying the gene of interest and is used to transfect insect cells. The need for transforming bacteria and isolating a large bacmid or co-transfecting a transfer vector and linear baculovirus DNA into insect cells is eliminated. As a result, the hands-on time is greatly reduced. Purified baculovirus can be isolated in less than one week.

Gateway linear DNA
BaculoDirect Linear DNA is designed for rapid cloning with a Gateway entry clone and subsequent expression in Sf9 or Sf21 insect cells. The Linear DNA features:

• Strong polyhedrin promoter for high-level expression
• R-sites for efficient recombination with any attL-flanked Gateway entry vector
• TK gene for negative selection using ganciclovir
• C-terminal V5-His tag (BaculoDirect™ C-Term Expression Kit) or N-terminal V5-His tag (BaculoDirect™ N-Term Expression Kit) for detection with anti-V5 antibody and purification with nickel-chelating resin
• TEV protease cleavage site for removal of the V5-His tag following purification (BaculoDirect N-Term Expression Kit)
• LacZ gene, to ensure a pure baculovirus stock is generated, which is replaced by your gene of interest after LR recombinant reaction

Additional materials required, available separately: Gateway entry clone.

A selection guide for choosing the most appropriate Gateway entry vector for your application can be found at: www.thermofisher.com/Gateway.

pMT/V5-His A, B, & C Drosophila Expression Vectors (Invitrogen™)

The DES®-Inducible Kit provides the expression vector pMT/V5-His for expression of recombinant proteins. This vector uses the Drosophila metallothionein gene promoter that is induced in S2 cells upon addition of copper sulfate or cadmium chloride to the culture medium.

The pMT/V5-His vector offers the following features:

• Small size (3.5 kb) to improve DNA yields and increase subcloning efficiency
• C-terminal V5 epitope tag for rapid detection with Invitrogen's Anti-V5 Antibody
• C-terminal 6xHis tag for simple purification of recombinant fusion proteins using nickel-chelating resin

To facilitate cloning, a set of three vectors-A, B, and C-is provided. Each vector has the multiple cloning site in a different reading frame relative to the coding sequence of the C-terminal tag.

Baculovirus Expression System with Gateway™ Technology (Invitrogen™)

The Baculovirus Expression System with Gateway® Technology is designed to create recombinant pFastBac™ plasmids containing the polyhedrin promoter. The Baculovirus Expression System with Gateway® Technology offers:

• Expression using the Bac-to-Bac® technology (1)
• Destination vectors carrying the polyhedrin promoter for production of native (pDEST™8), N-terminal histidine fusion (pDEST™10), and N-terminal GST fusion (pDEST™20) proteins
• pDEST™10 vector containing a TEV protease cleavage site for removal of the fusion tag after protein purification (2)

Gateway™ pDEST™20 Vector (Invitrogen™)

To fit all of your expression needs, Invitrogen offers state-of-the-art Gateway® destination vectors for expression in E. coli, insect, yeast, or mammalian cells, as well as for production of native protein or N- or C-terminal fusion proteins. All Gateway® destination vectors have attR sites for recombination with any attL-flanked fragment, regardless of whether it is an entry clone or an Ultimate™ RF Clone. The following table lists the wide range of destination vectors available.

Additional materials required, available separately: Gateway® entry clone, Gateway® LR Clonase® enzyme mix, and reaction buffer.

DES™-Inducible/Secreted Kit with pCoBlast (Invitrogen™)

The DES®-Inducible/Secreted Kit provides the vector pMT/BiP/V5-His for inducible, secreted expression of recombinant proteins and a choice of Blasticidin (pCoBlast) or hygromycin (pCoHygro) selection. This vector offers the inducible metallothionein promoter that is induced upon addition of copper sulfate or cadmium chloride. The N-terminal signal sequence from the insect BiP gene is provided to direct the recombinant fusion protein through the secretory pathway of S2 cells into the culture medium. The pMT/BiP/V5-His vector offers the following additional features:

• Small size (3.6 kb) to improve DNA yields and increase subcloning efficiency
• C-terminal V5 epitope tag for rapid detection with Invitrogens Anti-V5 Antibody
• C-terminal 6xHis tag for simple purification of recombinant fusion proteins using nickel-chelating resin

To facilitate cloning, a set of three vectors-A, B, and C-is provided. Each vector has the multiple cloning site in a different reading frame relative to the BiP signal sequence.

pMIB/V5-His Vector Kit (Invitrogen™)

pMIB/V5-His is a 3.6 kb vector designed for constitutive expression and secretion of recombinant proteins from Sf9, Sf21, and High Five™ insect cells. Secretion of proteins into serum-free medium simplifies purification, making it easier to harvest protein from cultured cells. pMIB/V5-His has several features to facilitate expression of recombinant proteins in insect cells including:

• The OpIE2 promoter for constitutive expression
• The honeybee melittin (HBM) secretion signal to allow secreted expression of your protein
• The Blasticidin resistance gene for rapid selection of stably transfected cell lines in two weeks
• C-terminal V5-epitope and polyhistidine tag for detection with Invitrogens Anti-V5 antibody and purification with nickel-chelating resin