Mr Lazarus Takawira
Biochemistry, Genetics and Microbiology
My Ph.D. research aims to decipher transcriptional regulatory networks involved in secondary cell wall biogenesis in Eucalyptus grandis. Understanding the transcriptional mechanisms that regulate secondary cell wall (SCW) development during wood formation in plants may enable strategic re-engineering of woody biomass traits. While extensive research has been devoted to model species such as Arabidopsis, much is yet to be learned about the nature of transcriptional networks underlying wood formation in hardwood trees such as Eucalyptus. We are exploring DNA Affinity Purification sequencing (DAP-seq) as a cost-effective high-throughput technique to interrogate protein-DNA interactions of Eucalyptus SCW-associated transcription factors (TFs).
This synthetic approach combines cell-free TF protein expression of NAC and MYB-Family TFs, immobilization of TF proteins, incubation with fragmented genomic DNA and wash steps before eluting bound DNA, in vitro. The eluted DNA will be sequenced, and generate reads, followed by read-mapping and the identification of positional weight matrices of TF binding sites using. Electrophoretic Mobility Shift Assay (EMSA_ assays will be carried out to validate these sequence logos and further characterize the cis-elements bound by these TFs and their potential target genes. Subsequently, the inference of SCW-associated transcriptional regulatory networks in Eucalyptus grandis will be done.