| Products/Services Used | Details | Operation |
|---|---|---|
| Gene Synthesis | A synthetic plant codon-optimized ORF was created for AY-WB SAP06 by GenScript Biotech (New Jersey, US) and subcloned by Gateway-based cloning into the entry vector pDONR207. The obtained donor vector was sequenced to confirm that the SAP06 ORF was correct and subsequently, a plant expression vector was generated by Gateway-based recombination. In this reaction, SAP06 was transferred into the destination vector pB7WG2 (Karimi et al., 2002), containing a CaMV 35S promoter-driven expression cassette. The obtained expression vector was transferred to Agrobacterium tumefaciens strain C58C1 and transformed into Arabidopsis Col0 by the floral dip method (Clough and Bent, 1998). Primary transformants were obtained upon selective germination of seeds harvested from the transformed plants on agar plates containing 10 mg/l phosphinotrycin (PPT). From these individual lines, six 3:1 segregating lines were selected based on selective germination on PPT medium, followed by selection of homozygous lines in the next generation. Subsequenty, qRT-PCR was used to select a transgenic line with a high ectopic expression level (SAP068) and a line with an intermediate expression level (SAP064). | Get A Quote |
Phytoplasmas are pathogenic bacteria that reprogram plant host development in order to attract their insect vectors to disseminate. Previous studies have characterized a few different phytoplasma effector proteins that supress specific transcription factors. However, these are only a small fraction of the potential effectors used by phytoplasmas, meaning that the molecular mechanisms through which phytoplasmas manipulate their hosts are largely uncharacterized. To obtain further insights into the phytoplasma infection mechanisms, we generated a protein-protein interaction network between a broad set of phytoplasma effectors and a large collection of Arabidopsis thaliana transcription factors and transcriptional... More
