DH10Bac cells harbor a baculovirus shuttle vector (bMON14272) and a helper plasmid (pMON7142), and are capable of supporting site-specific recombination between pFastBac and bMON14272 to generate high molecular weight bacmids that can then be amplified, purified, and used for insect cell transfection and subsequent gene expression.

• An E. coli host strain, DH10Bac™, that contains a bacmid and a helper plasmid, and allows generation of a recombinant bacmid following transposition of the pFastBac™ expression construct. • ExpiFectamine™ Sf Transfection Reagent for fast and efficient transfection of insect cells to generate recombinant baculovirus.

DH10Bac cells harbor a baculovirus shuttle vector (bMON14272) and a helper plasmid (pMON7142), and can generate high molecular weight bacmids that can then be amplified, purified, and used for insect cell transfection and subsequent gene expression.

Transformation of DH10Bac cells 1. Prepare LB agar plates containing 50 ug/mL kanamycin, 7 ug/mL gentamicin, 10 ug/mL tetracycline, 100 ug/mL Bluo-gal, and 40 ug/mL IPTG. 2. Add plasmid DNA to the competent cells and mix gently. 3. Incubate the mixture on ice for 30 minutes. 4. Heat-shock the mixture at 42°C for 45 seconds. Chill on ice for 2 ...

MAX Efficiency ™ DH10Bac Competent Cells are used to produce recombinant baculovirus molecules for the expression of eukaryotic proteins. The baculovirus may also be isolated and transfected into insect cells to produce infectious baculovirus particules.

• 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.

We describe here the creation of Bac-2-the-Future, a 2 nd generation Tn7-based system for recombinant baculovirus production which uses optimized expression vectors, new E. coli strains, and enhanced protocols to dramatically enhance the …

Transform the pFastBacTM construct into MAX Efficiency® DH10BacTM competent E. coli to generate a recombinant bacmid. Transfect the recombinant bacmid DNA into the insect cell line of choice to generate a recombinant baculovirus. Amplify and titer the baculoviral stock, and use this stock to infect insect cells to express your recombinant protein.

DH10Bac cells harbor a baculovirus shuttle vector (bMON14272) and a helper plasmid (pMON7142), and are capable of supporting site-specific recombination between pFastBac and bMON14272 to generate high molecular weight bacmids that can then be amplified, purified, and used for insect cell transfection and subsequent gene expression.

DH10Bac 细胞包含一种杆状病毒穿梭载体 (bMON14272) 和一种辅助性质粒 (pMON7142)，能够支持 pFastBac 与 bMON14272 的位点特异性重组以生成高分子量杆粒，然后对杆粒进行扩增、纯化并用于昆虫细胞转染和后续基因表达。