Today I completed Matt Nicholson's manuscript on the cloning and functional analysis of two indole-diterpene gene clusters from Penicillium. This work was started in 2006 so its rather sobering to look back at how rapidly the technology has changed - who would make a cosmid library these days when you can sequence the whole genome. However, we did use second generation Solexa technology in 2010 to sequence the cosmids. What will now make publishing the paper difficult is a recent publication from a group at Hokkaido University who in a tour de force have reconstituted penitrem biosynthesis in Aspergillus oryzae.
Yesterday I had very enjoyable discussions with two PhD students. Mariana has successfully developed the CRISPR/Cas9 technology for U. maydis. They have some very nice tools in U. maydis including a good selectable marker (cbx = carboxin) and an autonomously replicating vector. Once the selectable marker is removed the cells rapidly lose the plasmid and mutagenesis by CRISPR/Cas9 is shut down. They also have a nice selection system as the transition from yeast to filamentous growth generates a fuzzy phenotype on charcoal. CRISPR/Cas9 mutagenesis is a potentially powerful tool for silencing multiple genes such as those that comprise a gene family.
My second meeting was with Philipp who is studying an effector protein called Stp1 from effector cluster 10 of U. maydis. He has identified a putative plant interactor of this protein using Y2H and confirmed by co-IP. He is also making use of comparative genomics. We are fortunate to have such a good collection of fungal genome sequences from other Clavicipitaceae to inform our studies by comparative genomics - thanks to Chris Schardl.
Today there was a symposium for prospective students who want to study at the MPI for PhD. Each of them had to do a presentation of their MSc work. An interesting but highly selective way of recruiting students.
No comments:
Post a Comment