| Summary: | Progress Code: completed Statement: Values provided in temporal and spatial coverage are approximate only. Taken from the 2009-2010 Progress Report: Variations to work plan or objectives: No field work to collect samples was able to be performed, all activities to date have been laboratory based. 1. PCR analysis of PR in 65 Antarctic bacterial strains (obtained from previous AAD-supported activity) by marine science student Jaume Bibloni. Results to date indicate extensive presence of PR genes in Antarctic lake bacteria including undescribed species. PR presence in sea-ice bacteria was more restricted. Obvious strain-dependency occurs in PR distribution. Sequence data obtained for most PR positive strains. Limitations in the analysis include the need to use highly degenerate primer oligonucleotides (due to high level of variability PR sequences at the nucleotide level). Work is ongoing to retest several strains giving equivocal results in the PCR asays. 2. Proteomics analysis has been initiated (Clare Rutherford Honours student) for analysis of response of Psychroflexus torquis to light and dark conditions grown at 2 C (grown in a modified marine medium). The proteomics involves shotgun analysis using a 2-dimensional HPLC separation of trypsinised protein extracts (recovered using the QProteome proteoin extraction kit and extraction with membrane protein surfactant C7BzO, Sigma-Aldrich) coupled to nano-flow LTQ-Orbitrap mass spectrometry. This work is done in collaboration with Dr Edwin Lowe, Central Sciences Laboratory. The goal of this experiment is to determine whether the presence of light induces PR translation (abundance) and it cognate carotenoid hydroxylase as well as other changes to the proteome. Based on recent experiments on other bacteria (E. coli and L. monocytogenes) as much as 50% of the proteome can be recovered using this approach (termed "MudPit" - multidimensional protein information technology). This will be first time such ana analysis has been performed on a marine bacterial species. 3. ...
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