AB Vector - Plasmid DNA

  Plasmid DNA
 
  The first step in constructing any recombinant baculovirus is cloning the gene of interest into a plasmid DNA which is specifically designed for generation of recombinant baculoviruses. Such a plasmid is typically referred to as a "plasmid transfer vector" or a "donor plasmid" because it facilitates transfer of the gene of interest from the plasmid into baculovirus DNA.

Cloning a gene of interest is performed by routine techniques which most frequently involve digesting a plasmid with restriction endonucleases and subsequently ligating it with a PCR fragment encoding the gene of interest that was also digested using the same restriction endonucleases, the sites for which were introduced in the primers used for PCR. A gene of interest is inserted into multiple cloning sites (MCS) region of the plasmids under the control of the baculovirus polyhedrin promoter. Although alternative methods such as ligation free cloning and recombineering are possible, AB Vector plasmids are primarily designed for regular ligation based cloning and provide the polyhedrin promoter for efficient gene transcription. The polyhedrin promoter provides for the maximum level of protein expression that can be achieved using the baculovirus system. All AB Vector’s plasmids have Ampicillin as the resistance marker and can be easily transformed into common E.coli strains, such as XL10-Gold, DH5α, TOP10 etc. No special protocols are required for plasmid preparation. A miniprep preparation typically yields about 5-10 μg of plasmid DNA with an insert and this is more than sufficient for co-transfection and is of suitable quality.

AB Vector provides a large selection of plasmids to suite various protein expression and purification requirements as shown in the table below. A protein of interest can be expressed with either GST, His, GFP, MBP or dual His/ MBP, or FLAG/His tag or expressed without a tag using pVL1393 plasmid vector. Additionally, we offer vectors with a strong signal sequence (pAB-bee™, pAB-bee™-8xHis,pAB-bee-FH™ plasmid transfer vectors) to facilitate expression of secreted and membrane proteins.
 
  Relevant products
 
Plasmid transfer vectors                             Click on the product name for technical information!
Cat # Product Specification Unit size Price*
B1 pVL1393 High level of target protein expression 50 μl, 0.5 μg/μl $295.00
B2 pAcAB3 High level expression of up to 3 target proteins using a single recombinant
    baculovirus
50 μl, 0.5 μg/μl $395.00
B3 pAB-bee Provides target proteins with powerful honeybee melittin signal sequence to      facilitate translocation into the ER
Recommended for high level expression of secreted and transmembrane proteins
50 μl, 0.5 μg/μl $495.00
B3H pAB-bee™-8xHis Provides target proteins with powerful honeybee melittin signal sequence to      facilitate translocation into the ER
Recommended for high level expression of secreted and transmembrane proteins
N- or C-terminal 8xHis tag for affinity purification
50 μl, 0.5 μg/μl $495.00
B3FH pAB-bee™-FH Provides target proteins with powerful honeybee melittin signal sequence to      facilitate translocation into the ER
Recommended for high level expression of secreted and transmembrane proteins
C-terminal FLAG-8xHis tag for affinity purification and immonodetection
50 μl, 0.5 μg/μl $495.00
B4 pAB-6xHis™ High level expression of proteins with cleavable N-terminal 6xHis tag 50 μl, 0.5 μg/μl $295.00
B5 pAB-GST™ High level expression of proteins with cleavable N-terminal GST tag 50 μl, 0.5 μg/μl $295.00
B6 pAB-MBP™ High level expression of proteins with cleavable N-terminal MBP tag
MBP tag improves protein solubility and yield
50 μl, 0.5 μg/μl $395.00
B7 pAB-6xHis-MBP™ High level expression of proteins with cleavable N-terminal 6xHis MBP tag
High level expression of proteins with C-terminal MBP tag
MBP tag improves protein solubility and yield
50 μl, 0.5 μg/μl $495.00
B8 pVL-GFP High level expression of proteins with C-terminal GFP tag 50 μl, 0.5 μg/μl $395.00
B9 pVL-FH High level expression of proteins with C-terminal FLAG-8xHis tag 50 μl, 0.5 μg/μl $295.00
 
  *Go to PRODUCTS to order online. Alternatively, email an order as pdf attachment to info@abvector.com, or send it by Fax to 1-760-454-2465 or call 1-866-683-2867 to order by phone.

Click on PRODUCTS to view the entire list of recombinant baculovirus-related products.
 


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