Cloning of the glyceraldehyde 3-phosphate dehydrogenase gene of Flavescence dorée phytoplasma and development of serological and molecular tools for studying its expression.

The glyceraldehyde 3-phosphate dehydrogenase (gapA)
gene codes for a protein involved in the glycolytic
pathway and is commonly used in Real-Time RT-PCR
quantification studies as housekeeping gene. In this
work we cloned and sequenced the full-length gapA
gene from Flavescence dore´e phytoplasma (FDp).
A _35 kDa recombinant GapA protein was overexpressed
in Escherichia coli, purified and used as antigen
to raise anti-GapA rabbit polyclonal antibodies.
The antiserum detected the GapA protein by western
blot analysis of total protein extracts of FDp-infected
experimental host (Catharanthus roseus) and grapevine
plants collected in the field. We also developed an
FDp-specific gapA Taqman Real-Time RT-PCR assay
suitable for quantification overtime of gapA mRNA in
infected plants.