Day11.printed - What do you do to study this cell? 2...

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1 MBIO 3813-002 Day 11 Fri 9/16/11 Pic of Jeanine Rainbolt College of Ed Note an essential structural component (the wall and its bracing) and an external construction fence, which keeps some things out (recall the capsule). Not to anthropomorphize, but there are notable analogies to cell str. Topics (exam Monday) Characterizing Microbial Cell, cont’d – Capsule (mucoid polysacch or polypeptide) – Cytoplasmic membranes • Lipids in Archaea – Micromachines: ATPase, Flagellum – Cytoplasm Capsule: Klebsiella pneumoniae . (LeBoeffe lab manual) Possible roles for capsule : Hydration/resist drying Nutrients during starv’n Attachment to tissue or solid surfaces Keep some chemicals out Avoid phagocytosis Can be significant virulence factor in pathogens What to do to characterize cell • You have a new spp of microorganism in pure culture and 1 g of frozen cells. Almost nothing is known about it.
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Unformatted text preview: What do you do to study this cell? 2 characterizations Seq: 16S rRNA gene, or 16S rRNA More and more, seq some or all of genome Analyze seq for taxonomic purp Fractionate cell Break it apart and separate components by polarity, density, pI, size, chem affinity for separation matrices, polarity Solvent extraction, obtain Phospholipids Chemical hydrolysis of PL OH-, 100 C What do you get? Ester bond --> alcohol & acid Ether bond --> N.R. (try HI and 150 C) Ester (Alkali-labile) The ether links in Archaeal lipids give chemical stability to the membrane (well-needed for cells growing, e.g. at pH 2) The membrane-spanning tetraether stabilizes the membrane at high temps ( e.g. > 80 C). Cell can regulate what % of its lipids are tetraethers as reqd by temperature...
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Day11.printed - What do you do to study this cell? 2...

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