The DNA vaccines were designed as described before in Wagemakers et al. (29 (link)). From the B. burgdorferi N40 OspC gene sequence (NCBI reference DQ437463.1 and the respective tick salivary gland genes Salp15 (NCBI reference AAK97817.1), tHRF (NCBI reference DQ066335), TSLPI (NCBI reference AEE89466.1), and TIX-5 (NCBI reference AEE89467). The signal peptide (predicted by SignalP 4.0 web-based software, CBS, Lyngby, Denmark) was replaced with the human tissue plasminogen activator (hTPA) signal sequence (genbank AAA61213.1) (30 (link)). The resulting sequence was codon-optimized to mouse tRNA usage with Java Codon Adaptation tool (Braunschweig, Germany) (31 (link)). At the 5′ end a BamH1 and a Kozak sequence were added, and at the 3′ end a sequence encoding a double stop codon and a Xho1 were added. The insert was synthesized (BaseClear, Leiden, The Netherlands) and ligated into a BamH1/Xho1 restricted empty pVAX vector (Invitrogen, Carlsbad, CA, USA). The plasmid was amplified using a Nucleobond Xtra EF kit (Macherey-Nagel, Düren, Germany) and resuspended in DNase free water.
Bamh1 xho1 restricted empty pvax vector
Manufactured by Thermo Fisher Scientific
Sourced in Netherlands
The BamH1/Xho1 restricted empty pVAX vector is a laboratory tool used for cloning and vector construction purposes. It is a plasmid that has been digested with the restriction enzymes BamH1 and Xho1, creating compatible ends for insertion of DNA fragments. The pVAX vector backbone provides the necessary elements for plasmid replication and selection in bacterial hosts.
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Lab products found in correlation
2 protocols using bamh1 xho1 restricted empty pvax vector
1
Construction of DNA Vaccines for Lyme Disease
2
Design of pVAXhTPA-BB0405 DNA Vaccine
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The pVAXhTPA-BB0405 DNA vaccine was designed as described before in Wagemakers et al.10 (link). In the BB0405 gene sequence of B. burgdorferi B31 (NCBI Reference Sequence: NC_001318.1) the 18aa signal sequence (predicted by SignalP 4.0 web-based software, CBS, Lyngby, Denmark) was replaced with the human tissue plasminogen activator (hTPA) signal sequence (genbank AAA61213.1)26 (link). The resulting sequence was codon-optimized to mouse tRNA usage with Java Codon Adaptation tool (Braunschweig, Germany)27 (link). At the 5′ end a BamH1 and a Kozak sequence were added, and at the 3′ end a sequence encoding a double stop codon and a Xho1 were added. The insert was synthesized (BaseClear, Leiden, The Netherlands) and ligated into a BamH1/Xho1 restricted empty pVAX vector (Invitrogen, Carlsbad, CA, USA). The plasmid was amplified using a Nucleobond Xtra EF kit (Macherey-Nagel, Düren, Germany) and resuspended in DNase free water.
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