Heungbok Kim

Activities, Skills and Qualification: . Member, TB Structural Genomics Consortium (2007~Present) . Member, International Structural Genomics Organization (2007~Present) . Bioscience Worker Safety and Security Team (BWSST, 2010~2012) . Chemistry, Life & Earth Sciences (CLES) tech of the year selection committee (2010), . Cytometry, Spectroscopy, and Instrumentation Team (CSIT) . Designated Procurement Representative (DPR), Admin work, Purchasing Items . Supervising students (training, instructing, and assigning student work) . Optimizing protocols for biological experiment ->MembraneProtein:expression, cell breakage and membrane fractionation, solubilization and purification in the presence of detergents- purified & crystallized AcrB, OprA, BpeF, AmrA, and AmrB (efflux pump) membrane proteins ->Molecular Biology: Making competent cells, Designing primers, Cloning, Protein Expression & Purification-LIC (Ligation Independent Clone) Complex, pCHis-Sub clone, pET/pETM/pMCSG clone, Surface Mutation, MBP fusion TB proteins, Fab Chaperone, Complex proteins, Sel-Met proteins through column chromatography (Protein A column, S protein column, His-tag affinity chromatography, Gel filtration chromatography, or Ion Exchange chromatography etc.), western blot, etc. ->Crystallization & data collection: Screening, optimizing, seeding, & harvesting crystals, collecting crystal data, accessing computer to data processing, refining and analyzing crystal data etc. Structure Solved (6 Mtb proteins - Rv2523c, Rv3275, Rv0927, Rv1122, Rv0155, & Rv2780), ->Dye-ligand affinity chromatography (DLAC) technique development – ID of hypothetical protein function with ligands, drug target screening of membrane and cytosolic fractions by DLAC ->Various assays: Enzymatic assay, Enzymatic kinase Assay, drug activity/inhibition assay, ligand/drug binding assay, MIC(the minimal inhibitory concentration) of drug compound, etc… ->Mass Spec: 1D and 2D gel running with immobiline dry strip, In Gel digestion for MALDI, purification & concentration of peptides isolated from in-gel digestion of protein bands by zip-tip, MS data analysis ->A multiplex oligonucleotide ligation-polymerase chain reaction (MOL-PCR) Based Assays for the Detection of Respiratory Pathogens: A multiplex assay for sequence detection/genotyping using Luminex Instrument - Kinase Reaction, Moligo Mix, Oligo Ligation, Symmetric PCR, Asymmetric PCR, Hybridization Reaction, Luminex Instrument operation, Conjugation of Address tag to Luminex Bead ->Influenza virus: infecting and harvesting the virus from the infected mammalian cells; influenza plaque assay, plaque fixation and staining with crystal violet; growing, passaging, freezing mammalian cells for the surface receptors (HA & NA) on the influenza viruse; labeling protein (SNA) with alexa 488-TFP; purification of labeled protein; conjugation of MDCK cells with labeled protein; titration of conjugated MDCK cells with SNA-TFP, Autophage [work on solving structures of NS1, CrkL-NS1complex, tri-complex (hp85β-CrkLNS1), and on drug screening assay with NS1] ->AIV Surveillance Project (An Early Detection System for Asian H5N1 Highly Pathogenic Avian Influenza in Wild Migratory Birds U.S. Interagency Strategic Plan, Evaluating Transmission Pathways of Avian Influenza (H5N1) by Domestic and Wild Birds in Central Africa): Set up BSL 2 lab, Tested ~200 virus spiked animal fecal samples (chicken, dog, and cat) to compare Ambion viral kit (Kingfisher 24well plate) with standard Qiagen RNAese kit for RNA extraction efficiency, Screened ~700 Avian fecal/cloacal swab samples in 100% ethanol with Kingfisher 96 robot (RNA purification) and Real time RT-PCR: The complete protocol was transferred to UCLA ->Yeast cell Growing(AH109), Single/Co-transformation of plasmids into yeast, Rapid Transformation of plasmid into yeast, Making a master plate & Replication (X-α-gal), Mammalian two-hybrid system: growing and splitting mammalian cells (#293 & GB133), Co-transfecting plasmids into mammalian cells, and harvesting the co-transfected mammalian cells ->Flow Cytometry: Titration of spherotech beads, conjugation of protein to beads, fluorescent staining (CBQCA) on microspheres, Block on Luminex beads, CBQCA binding to avidin microsphere, kinetic reaction of CBQCA etc. Enzymatic Assay of aldehyde dehydrogenases, Screen recombinant TB proteins for binding to Affigel Blue ->Microsphere based protease assays: Assays using Lethal factor (LF cleavage of protein such as Mek2Gfp – bead based, solution based), Lethal Factor cleavage of phage bound to magnetic/agarose bead (avidin microspheres), etc. ->Phage Display: phage titration, Phage Amplification, Construction of consensus library, electroporation, phage infection, etc.