Gibco™

Bac-to-Bac™ HBM 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 (DH10Bac™) rather than homologous recombination in insect cells. The new pFastBac™ HBM TOPO™ vector enables secreted protein expression because it has the honeybee melittin (HBM) secretion signal. The new vector should be strongly considered for the expression of glycoproteins. Glycoproteins cannot be glycosylated in the absence of any secretion signal. In contrast to glycoproteins secreted from mammalian cells, glycoproteins secreted from baculovirus can be easily de-glycosylated in vitro. This is a very important feature in order to crystallize proteins!

The pFastBac™ HBM TOPO™ vector has also a C-terminal His-Tag with a TEV cleavage signal to enable easy purification of Histidine fusion proteins on nickel-chelating resins (ProBond™ Purification System) and native proteins with the aid of AcTev™ protease.
The vector uses the strong polyhedrin promoter to generate high levels of expression in a variety of insect cell lines such as Sf9, Mimic™ Sf9, Sf21, and High Five™ cells in Sf-900 II & Sf-900™ III media from Gibco.

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™ vectors, 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.

사양

세포주High Five™, Sf21, Sf9

세포 유형Insect Cell

클로닝 방법Blunt TOPO

분열TEV Protease Recognition Site

형식Kit

발현 메커니즘Cell-Based Expression

발현 시스템Baculovirus

용도(애플리케이션)Protein Expression

고처리량 호환성High-throughput Compatible, Not High-throughput Compatible (Manual)

주요 기능Secreted Expression

반응 수20

제품라인Bac-to-Bac™, TOPO™

제품 유형Cloning Kit

프로모터Polyhedrin

단백질 태그HBM (Honeybee Melittin), His Tag (6x)

선택 제제(진핵)None

벡터pFastBac/HBM-TOPO

Unit SizeEach