Articles by Sandra Vomund in JoVE
Generation of Topically Transgenic Rats by In utero Electroporation and In vivo Bioluminescence Screening Sandra Vomund1, Tamar Sapir2, Orly Reiner2, Maria A. de Souza Silva3, Carsten Korth1 1Department of Neuropathology, Medical School Düsseldorf, 2Department of Molecular Genetics, Weizmann Institute for Science, 3Center of Behavioral Neuroscience, University of Düsseldorf Genes can be manipulated during development of the cortex or the hippocampus of the rat via in utero electroporation (IUE) at E16, to enable fast and targeted modifications in neuronal connectivity for later studies of behavior or neuropathology in adult animals. Postnatal in vivo imaging for control of IUE success is performed by bioluminescence of activating co-transfected luciferase.
Other articles by Sandra Vomund on PubMed
Rational Design of a Minimal and Highly Enriched CYP102A1 Mutant Library with Improved Regio-, Stereo- and Chemoselectivity Chembiochem : a European Journal of Chemical Biology. Mar, 2009 | Pubmed ID: 19222039 A minimal CYP102A1 mutant library of only 24 variants plus wild type was constructed by combining five hydrophobic amino acids (alanine, valine, phenylalanine, leucine and isoleucine) in two positions. Both positions are located close to the centre of the haem group. The first, position 87, has been shown to mediate substrate specificity and regioselectivity in CYP102A1. The second hotspot, position 328, was predicted to interact with all substrates during oxidation and has previously been identified by systematic analysis of 31 crystal structures and 6300 sequences of cytochrome P450 monooxygenases. By systematically altering the size of the side chains, a broad range of binding site shapes was generated. All variants were functionally expressed in E. coli. The library was screened with four terpene substrates geranylacetone, nerylacetone, (4R)-limonene and (+)-valencene. Only three variants showed no activity towards all four terpenes, while eleven variants demonstrated either a strong shift or improved regio- or stereoselectivity during oxidation of at least one substrate as compared to CYP102A1 wild type.