Construction of DNA Vaccines for Lyme Disease

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.

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Klouwens M.J., Trentelman J.J., Wagemakers A., Ersoz J.I., Bins A.D, & Hovius J.W. (2021). Tick-Tattoo: DNA Vaccination Against B. burgdorferi or Ixodes scapularis Tick Proteins. Frontiers in Immunology, 12, 615011.

Publication 2021

BamH1/Xho1 restricted empty pVAX vector Nucleobond Xtra EF kit

Adaptation Codon Dna vaccines Dnase Genes Human Mouse Ospc Plasmid Salivary gland Signal sequence Stop codon Tick Tissue plasminogen activator Trna Vector

Corresponding Organization : University of Amsterdam

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independent variables

The DNA vaccines were designed as described before in Wagemakers et al. (29 (link)).
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).

dependent variables

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control variables

Not explicitly mentioned.

controls

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