Expression of the tuf Gene of "Candidatus Phytoplasma mali" in Escherichia coli

O. Viczián, E. Mergenthaler, S. Süle

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

The tuf gene of "Candidatus Phytoplasma mali", the causal agent of apple proliferation was PCR cloned in an expression vector and expressed in Escherichia coli. First, phytoplasma DNA extracted from periwinkle was amplified using primers designed on the basis of the tuf gene and the PCR product was cloned into pGEM-T (Promega). In the next step specific primers were constructed containing some plasmid sequences and restriction enzyme sites. With this primers the sequence in pGEM-T was amplified, the product was digested with restriction enzymes, and inserted into the pQE40 expression vector (Qiagen). In this plasmid the tuf gene was fused to the 6xHIS tag, and DHFR. The production of 6xHIS-DHFR-Tu fusion protein protein was induced with IPTG and expressed in E. coli M15. The new fusion protein was found in the insoluble fraction of the bacterium. The identity of the protein was verified with polyacrylamid gel-electrophoresis and Western blot analysis using antiserum raised against the 6xHIStag of the fusion protein.

Original languageEnglish
Pages (from-to)225-231
Number of pages7
JournalActa Phytopathologica et Entomologica Hungarica
Volume40
Issue number3-4
DOIs
Publication statusPublished - Dec 1 2005

Keywords

  • Apple proliferation
  • Phytoplasma
  • pQE40
  • tuf gene

ASJC Scopus subject areas

  • Plant Science
  • Insect Science

Fingerprint Dive into the research topics of 'Expression of the tuf Gene of "Candidatus Phytoplasma mali" in Escherichia coli'. Together they form a unique fingerprint.

  • Cite this