VE-BEVS Transfer Vectors Resource Page

pAcVE1 Transfer Vector Instructions

 Contents

The plasmid DNA was prepared on a silicon bead matrix and dissolved in TE buffer (10mM Tris-HCl, pH7.5; 1 mM EDTA). pAcVE1 baculovirus transfer vector is provided at 10 µg in 20 µl.

Storage

Transfer vector should be placed at -20°C for long- term storage.

Handling

For expression of recombinant protein under the polyhedrin promoter, the gene of interest should be ligated into an appropriate restriction site in the multiple cloning site of the pAcVE1 vector (see below).  Transform the recombinant pAcVE1 transfer vector in E. coli cells (Top10 or any other suitable strain), propagate the cells under ampicillin selection and purify the recombinant plasmid using standard plasmid purification protocols.  For construction of recombinant AcMNPV virus perform a co-transfection of the purified recombinant pAcVE1 vector with linearized baculovirus DNA suitable for homologous recombination in insect cells.

pAcVE1 Vector Map and Multiple Cloning Site

Click here to view the "pAcVE1 Vector Map and Multiple Cloning Site" image.

Unique Restriction Sites

AgeI   AlwNI   ApaI   AscI   AvrII   BglII   BsaAI   Bsp120I   BssHII   BstAPI   BstXI   ClaI   DraII   EagI   EcoNI   EcoRI   FspAI   MscI   NaeI   NdeI   Ngo   MIV   NheI   NotI   NruI   PacI   PfoI   SbfI   SgrAI   SmaI   SnaBI   SpeI   StyI   SwaI   XhoI   XmaI

Absent Restriction Sites

AflII   BclI   BlpI   BsiWI   BstEII   BstZ17I   Bsu36I   DraIII   FseI   MboI   NcoI   PflMI   PmeI   PmlI   PpuMI   PshAI   RsrII   SacII   SanDI   SexAI   SfiI   SgfI   SrfI   Tth111I   XcmI

 

Click here to view the "Yellow Fluorescent Protein expression is enhanced when pAcVE1 is used to generate a vankyrin-enhanced baculovirus" image.

THIS PRODUCT IS INTENDED FOR RESEARCH PURPOSES ONLY

CAUTION: Not intended for human or animal diagnostic or therapeutic uses.

 

Limited Warranty

ParaTechs warrants that, at the time of shipment, the Product will conform to the specifications that accompany the Product. This warranty limits ParaTechs liability to replacement of the Product. PARATECHS MAKES NO OTHER WARRANTIES, EXPRESSED OR IMPLIED, WITH RESPECT TO THE PRODUCT; INCLUDING ANY WARRANTIES OF MERCHANTABILITY OR FITNESS FOR ANY PARTICULAR PURPOSE OR THAT THE PRODUCT DOES NOT INFRINGE ANY PROPRIETARY RIGHTS OF ANY THIRD PARTY.

 

pAcVE.02 Transfer Vector Instructions

 Contents

The plasmid DNA is dissolved in TE buffer (10mM Tris-HCl, 1 mM EDTA pH7.5). pAcVE.02 baculovirus transfer vector is provided at 10µg in 50µl.

Storage

The transfer vector should be placed at -20º C for long-term storage.

Handling

For expression of recombinant protein under the polyhedrin promoter, the gene of interest should be ligated into an appropriate restriction site in the multiple cloning site of the pAcVE.02 vector and should be in frame with the N-terminal His-tag (see diagram). Transform the recombinant pAcVE.02 transfer vector in E. coli cells (DH5α, Top10 or any other suitable strain), propagate the cells under ampicillin selection and purify the recombinant plasmid using standard plasmid purification protocols. For construction of recombinant AcMNPV perform a co-transfection of the purified recombinant pAcVE.02 vector with linearized baculovirus DNA suitable for homologous recombination in insect cells.

pAcVE.02 Vector Map and Multiple Cloning Site

Click here to view the "pAcVE.02 Vector Map and Multiple Cloning Site" image.

Unique Restriction Sites

AgeI   AleI   AlwNI   ApaI   AvaI   AvrII   BamHI   BbvCI   BclI   BmgBI   BmtI   BplI   BpmI   Bpu10I   BsaAI   BsaBI   BseRI   BseYI   BspMI   BstAPI   BstBI   BstXI   EagI   Eco53kI   EcoNI   EcoRI   HindIII   HpaI   KpnI   NaeI   NdeI   NgoMIV   NheI   NotI   NruI   PciI   PfoI   PmeI   PpuMI   PspOMI   SacI   SapI   SbfI   SgrAI   SmaI   SnaBI   SpeI   SphI   StuI   StyI   SwaI   XbaI   XmaI

Absent Restriction Sites

AscI   AsiSI   BglII   BlpI   BsiWI   BspEI   BssHII   BstEII   BstZ17I   Bsu36I   BtgI   DraIII   FseI   FspAI   MscI   NcoI   PflMI   PmlI   PshAI   RsrII   SacII   SanDI   SexAI   SfiI   Tth111I   XcmI   XhoI

 

Click here to view the "Secreted alkaline phosphatase (SEAP) activity is enhanced when expressed from pAcVE.01" image.

THIS PRODUCT IS INTENDED FOR RESEARCH PURPOSES ONLY

CAUTION: Not intended for human or animal diagnostic or therapeutic uses.

 

Limited Warranty

ParaTechs warrants that, at the time of shipment, the Product will conform to the specifications that accompany the Product. This warranty limits ParaTechs liability to replacement of the Product. PARATECHS MAKES NO OTHER WARRANTIES, EXPRESS OR IMPLIED, WITH RESPECT TO THE PRODUCT; INCLUDING ANY WARRANTIES OF MERCHANTABILITY OR FITNESS FOR ANY PARTICULAR PURPOSE OR THAT THE PRODUCT DOES NOT INFRINGE ANY PROPRIETARY RIGHTS OF ANY THIRD PARTY.

 

pAcVE.03 Transfer Vector Instructions

 Contents

The plasmid DNA is dissolved in TE buffer (10mM Tris-HCl, 1 mM EDTA pH7.5). pAcVE.03 baculovirus transfer vector is provided at 10 µg in 50 µl.

Storage

The transfer vector should be placed at -20º C for long-term storage.

Handling

For expression of recombinant protein under the polyhedrin promoter, the gene of interest should be ligated into an appropriate restriction site in the multiple cloning site of the pAcVE.03 vector and should be in frame with the honey bee melittin signal sequence and the C-terminal 6xHis-tag if desired (see diagram and table). In that instance the stop codon needs to be omitted. Transform the recombinant pAcVE.03 transfer vector in E. coli cells (DH5α, Top10 or any other suitable strain), propagate the cells under ampicillin selection and purify the recombinant plasmid using standard plasmid purification protocols. For construction of recombinant AcMNPV perform a co-transfection of the purified recombinant pAcVE.03 vector with linearized baculovirus DNA suitable for homologous recombination in insect cells.

pAcVE.03 Vector Map and Multiple Cloning Site

Click here to view the "pAcVE.03 Vector Map and Multiple Cloning Site" image.

Restriction Enzymes Suitable for In-Frame Cloning

Click here to view the "Restriction Enzymes Suitable for In-Frame Cloning" image.

Unique Restriction Sites

AgeI   AleI   AlwNI   ApaI   AvrII   BamHI   BbvCI   BclI   BglII   BmgBI   BplI   BpmI   Bpu10I   BsaAI   BsaBI   BseRI   BseYI   BstAPI   BstXI   BstZ17I   EagI   Eco53kI   EcoNI   EcoRI   HindIII   HpaI   KpnI   NaeI   NcoI   NdeI   NgoMIV   NotI   NruI   PciI   PfoI   PpuMI   PspOMI   SacI   SacII   SapI   SbfI   SgrAI   SmaI   SnaBI   SpeI   SphI   StuI   SwaI   XbaI   XhoI   XmaI

Absent Restriction Sites

AscI   AsiSI   BlpI   BmtI   BsiWI   BspEI   BspMI   BssHII   BstEII   Bsu36I   DraIII   FseI   FspAI   MscI   NheI   PflMI   PmeI   PmlI   PshAI   RsrII   SanDI   SexAI   SfiI   Tth111I   XcmI

 

Click here to view the "Erythropoietin (EPO) production is enhanced when expressed from pAcVE.03" image.

THIS PRODUCT IS INTENDED FOR RESEARCH PURPOSES ONLY

CAUTION: Not intended for human or animal diagnostic or therapeutic uses.

 

Limited Warranty

ParaTechs warrants that, at the time of shipment, the Product will conform to the specifications that accompany the Product. This warranty limits ParaTechs liability to replacement of the Product. PARATECHS MAKES NO OTHER WARRANTIES, EXPRESS OR IMPLIED, WITH RESPECT TO THE PRODUCT; INCLUDING ANY WARRANTIES OF MERCHANTABILITY OR FITNESS FOR ANY PARTICULAR PURPOSE OR THAT THE PRODUCT DOES NOT INFRINGE ANY PROPRIETARY RIGHTS OF ANY THIRD PARTY.

 

VE-BEVS Publications

 

Fath-Goodin A, Kroemer JA, Webb BA (2009). The Campoletis sonorensis ichnovirus vankyrin protein P-vank-1 inhibits apoptosis in insect Sf9 cells. Insect Mol Biol. 18(4):497-506. PMID: 19453763. https://pubmed.ncbi.nlm.nih.gov/19453763/

Fath-Goodin A, Kroemer J, Martin S, Reeves K, Webb BA (2006). Polydnavirus genes that enhance the baculovirus expression vector system. Adv Virus Res. 68:75-90. Review. PMID: 16997009. https://pubmed.ncbi.nlm.nih.gov/16997009/

Kroemer JA, Webb BA (2006). Divergences in protein activity and cellular localization within the Campoletis sonorensis Ichnovirus Vankyrin family. J Virol. 80(24): 12219-28. PMID: 1700565. https://pubmed.ncbi.nlm.nih.gov/17005654/

 

VE-BEVS Presentations

 

Steele K, oral presentation entitled “Expression of mammalian glycoproteins and other difficult to express proteins with the vankyrin-enhanced baculovirus expression system.” Presented during PEGS: The Essential Protein Engineering Summit, Boston, MA, May 6, 2014. [PDF]

Steele K, poster presentation entitled “End the poor protein yield crisis by delaying cell lysis: new baculovirus transfer vectors encoding the anti-apoptotic vankyrin protein.” Presented during the International Society for BioProcess Technology, 4th Spring meeting, Washington D.C., March 12, 2014

Oral presentation entitled “Function and Versatility of Vankyrin Gene-Mediated Enhancement of Baculovirus Expression Vector Protein Production” at ISBiotech 2nd Annual Meeting “Baculovirus Expression Technology”, Rosslyn, VA, April 2-5, 2012

Poster presentation entitled “Log-Scale Improvement in Protein Yield with Vankyrin-Enhanced Transfer Vector” at the Cambridge Healthtech Institute “Baculovirus Technology” conference, Boston, MA, August 22-23, 2011 [PDF]

Oral presentation entitled “Expanding the Utility of Viral Ankyrin Genes in Baculoviruses” at the Wilbio 12th International Conference on Baculovirus and Insect Cell Culture, San Antonia, TX, February 2-4, 2009

Oral presentation entitled “Improved Recombinant Protein Production by ParaTechs’ Vankyrin-Enhanced Baculovirus Expression Technology” ” at the Cambridge Healthtech Institute “Baculovirus Technology” conference, Boston, MA, August 2008

Poster presentation entitled “Enhanced Recombinant Protein Production by ParaTechs’ Vankyrin Enhanced Baculovirus Expression Technology” at the Wilbio 11th International Conference on Baculovirus and Insect Cell Culture, Seattle, WA, February 25-27, 2008 [PDF]

 

VE-BEVS Licensing

 

The VE-BEVS Cell Line (“Product”) and VE-BEVS Transfer Vectors (“Product”) were developed in collaboration by scientists at ParaTechs and the University of Kentucky Lexington for expression of recombinant proteins. One or more patents or patent applications owned by the University of Kentucky Lexington cover components of the Product.

ParaTechs Corporation has an exclusive license to sell the Product to scientists for academic research or one year commercial evaluation only, under the terms described below. Use of the Product for any Commercial Purpose (as defined below) other than evaluation requires the user to obtain a commercial license as detailed below. Before using the Product, please read the terms and conditions set forth below. Your use of the Product shall constitute acknowledgement and acceptance of these terms and conditions. If you do not wish to use the Product pursuant to these terms and conditions, please contact ParaTechs Technical Service Department to return the unused and unopened Product for full credit.

ParaTechs grants the purchaser a non-exclusive license to use the enclosed Product for academic research or for commercial evaluation purposes only. The Product is being transferred to you in furtherance of, and reliance on, such license. You may not use the Product, or the materials contained therein, for any Commercial Purpose without a license for such purpose from ParaTechs Corporation.

Commercial Purpose include any use of Product in a Commercial Product, the manufacture of a Commercial Product, any resale of the Product, any use (other than evaluation) of Product to facilitate or advance research or development of a Commercial Product, and any use (other than evaluation) of the Product to facilitate or advance any research or development program the results of which will be applied to the development of Commercial Products.

“Commercial Product” means any product intended for commercial use.

Access to the Product must be limited solely to those officers, employees and students of your entity who need access to perform the aforementioned research or evaluation. Each such officer, employee and student must be informed of these terms and conditions and agree in writing, to be bound by same. You may not distribute the Product to others. You may not transfer modified, altered, or original material from the Product to a third party without written notification to and written approval from ParaTechs.

Inquiries for commercial use should be directed to agoodin@paratechs.com.

Patent information:
VE-BEVS Cell Line United States Patent 7,842,493 [PDF]
VE-BEVS Transfer Vector United States Patent 7,629,160 [PDF]

 

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