form
liquid
manufacturer/tradename
Novagen®
storage condition
OK to freeze
avoid repeated freeze/thaw cycles
storage temp.
−20°C
General description
The pBACsurf-1 transfer plasmid is designed for in-frame insertion of target genes between the gp64 signal sequence and the mature protein coding sequence (under the control of the polh promoter). Expressed fusion proteins, including glycoproteins, are secreted and incorporated onto the virion surface, anchored by the transmembrane domain of gp64 (1). Target proteins are displayed at the poles of the virion, where they appear to be localized as hetero-oligomers with the wild-type gp64 also encoded by the virus (1).
For virus display, inserts must lack an internal stop codon and maintain the appropriate open reading frame. Inserts in-frame with the N-terminal gp64 signal sequence and carrying an internal stop codon can produce target proteins that may be secreted from the recombinant virus-infected cell, provided they lack a membrane-spanning sequence. Alternatively, target proteins may be displayed on the cell surface and on budded virions if they remain in-frame with the downstream gp64 gene.
The plasmid is compatible with BACVECTOR-1000, -2000 or -3000 Triple Cut Virus DNA for low background transfection and efficient utilization of the polh promoter. The cloning/expression region of the coding strand transcribed from the polh promoter is shown on the vector map (TB148). The f1 origin is oriented so that infection with helper phage will produce virions containing single stranded DNA that corresponds to the non-coding strand. Single stranded sequencing of phage-derived DNA can be performed using the gp64 signal primer.
For virus display, inserts must lack an internal stop codon and maintain the appropriate open reading frame. Inserts in-frame with the N-terminal gp64 signal sequence and carrying an internal stop codon can produce target proteins that may be secreted from the recombinant virus-infected cell, provided they lack a membrane-spanning sequence. Alternatively, target proteins may be displayed on the cell surface and on budded virions if they remain in-frame with the downstream gp64 gene.
The plasmid is compatible with BACVECTOR-1000, -2000 or -3000 Triple Cut Virus DNA for low background transfection and efficient utilization of the polh promoter. The cloning/expression region of the coding strand transcribed from the polh promoter is shown on the vector map (TB148). The f1 origin is oriented so that infection with helper phage will produce virions containing single stranded DNA that corresponds to the non-coding strand. Single stranded sequencing of phage-derived DNA can be performed using the gp64 signal primer.
Warning
Toxicity: Standard Handling (A)
Other Notes
1. Boublik, Y., Di Bonito, P., and Jones, I.M. (1995) Bio/Technology 13, 1079–1084.
Legal Information
NOVAGEN is a registered trademark of Merck KGaA, Darmstadt, Germany
Storage Class
10 - Combustible liquids
wgk_germany
WGK 1
Certificates of Analysis (COA)
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