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Development Of Rapid, Homogeneous Assay For Investigating Isopeptide Bond Formation Using Fluorescence Polarization/Depolarization Measurements, Samuel Patricc Kasson Aug 2018

Development Of Rapid, Homogeneous Assay For Investigating Isopeptide Bond Formation Using Fluorescence Polarization/Depolarization Measurements, Samuel Patricc Kasson

MSU Graduate Theses

Autocatalytic intramolecular isopeptide bonds have been found in nature in certain gram-positive bacterial pilus structures. Recently, splitting of these domains that are capable of autocatalytic intramolecular isopeptide bond formation have been applied to create stable, selective, bio-orthogonal Catcher/Tag systems. The CnaB2 domain found in the FbaB pilus structure of Streptococcus pyogenes, has yielded the Catcher/Tag, Protein/Peptide systems termed SpyCatchter and SpyTag. Recent study has focused on tag optimization, stability and bio-orthogonality evaluation, along with applications in bioconjugation. I have recombinantly expressed SpyCatcher and SpyTag-fused proteins in E.coli, and conjugated them to fluorescent probes in order for use in fluorescence polarization/depolarization …


Assessment Of Resistance To Botrytis Cinerea In Arabidopsis Expressing Grapevine Sts Genes And Analysis Of New Grapevine Vein Clearing Virus Isolates, Li Su May 2018

Assessment Of Resistance To Botrytis Cinerea In Arabidopsis Expressing Grapevine Sts Genes And Analysis Of New Grapevine Vein Clearing Virus Isolates, Li Su

MSU Graduate Theses

Grapevine stilbene synthase gene (STS) family is unusually large with more than 32 members of full-length genes. Resveratrol and piceid are synthesized in VaSTS7-, VvSTS22-, VaSTS22-transgenic Arabidopsis. To study the function of these STS genes in the resistance against Botrytis cinerea in vivo, lesion sizes on these STS transgenic Arabidopsis were compared to that on non-transgenic control Col-0, at 96 hours post inoculation with B. cinerea spores at 0, 200, or 1,000 spores/μl. In each experiment there were three biological replicates. Three lesion sizes from each replicate were averaged. The experiment was repeated three …