Gibco™

Bac-to-Bac™ C-His TOPO™ Cloning Kit

Bac-to-Bac™ Baculovirus Expression is an efficient method for producing baculovirus for high-level protein expression in insect cells. It relies on자세히 알아보기

Bac-to-Bac™ Baculovirus Expression is an efficient method for producing baculovirus for high-level protein expression in insect cells. It relies on generation of recombinant virus by site-specific transposition in E. coli rather than homologous recombination in insect cells. New: pFastBacTOPO™ vectors!

Reliable and fast protein expression
The Bac-to-Bac™ Baculovirus Expression System is faster and easier than traditional baculovirus expression, and it maintains high levels of protein expression. Bac-to-Bac™ relies on generation of recombinant baculovirus by site-specific transposition in E. coli rather than homologous recombination in insect cells. The Bac-to-Bac™ Baculvirus Expression System is highly regarded in academic Literature. Approximately 100 citations every year since 2000!

Traditional baculovirus systems require purification and amplification of an initial low-titer viral supernatant. This requires a time-consuming plaque assay. Bac-to-Bac™s pFastBac™ vector, however, recombines with the parent bacmid in DH10Bac™ E. coli competent cells to form an expression bacmid. Transfect the bacmid into insect cells for fast production of a high titer of pure recombinant baculovirus particles in the very first transfection. Youll save weeks of precious time. Collect a pure P2 baculovirus stock on Day 10 without the necessity of tedious plaques assay (See Figure 1)

High expression, easy screening
pFastBac™ uses the strong polyhedrin promoter to generate high levels of expression in a variety of insect cell lines such as Sf9, Sf21, and High Five™ cells.

New additions:
Bac-to-Bac™`s pFastBac vectors are now available as pFastBacTOPO™ vectors too. pFastBacTOPO™ vectors are Blunt™ TOPO™ vectors and are supplied with Mach1™-T1R E. coli for easy cloning and pFastBac⁄GOI propagation. The new vectors enable you to:

• Amplify your gene of interest (GOI) with a PCR enzyme of highest fidelity such as Accuprime™ Pfx SuperMix.

• TOPO™ clone the blunt-end PCR product into the new vector in only 5 minutes!

• Visualize colonies 8 hours after plating on ampicillin selective plates because Mach1™-T1R E. coli cells have a faster doubling time compared to other standard cloning strains.

• With the Bac-to-Bac™ C-His TOPO™ cloning or expression kit, you can produce a C-terminal His-fusion protein with a TEV cleavage site to purify with nickel-chelating resins (Invitrogens ProBond™ Purification System) and generate a native protein with the aid of Invitrogens popular AcTEV™ Protease. This is the Invitrogen`s only vector that contains a C-terminal TEV cleavage site!

The new vectors combine the TOPO™ cloning technology with the highly regarded Bac-to-Bac™ baculovirus expression features to enable easy cloning and high-level protein expression.
The pFastBacTOPO™ vectors are offered as cloning and expression kits.

사양

항생제 내성 박테리아Ampicillin (AmpR), Gentamicin (GmR)

세포주High Five™, Sf21, Sf9

세포 유형Insect Cell

클로닝 방법Blunt TOPO

발현 메커니즘Insect Cell Expression

발현 시스템Baculovirus

용도(애플리케이션)Protein Expression, Viral Vaccine Production

반응 수20 Reactions

제품라인Bac-to-Bac™, TOPO™

제품 유형Cloning Kit

프로모터Polyhedrin

단백질 태그His Tag (6x)

수량20 Reactions

선택 제제(진핵)Ampicillin, Gentamicin

벡터pFastBac/CT-TOPO

Unit SizeEach