• Name: ANASTASSIA M. SAVILOVA (Bushueva)
  
• University: Moscow State University, Biological faculty, Department of Molecular Biology.
  
• Address: poste restante (do vostrebovaniya) Porechnaya str., 109469, Moscow, Russia.
  
• Telephone (work): 007 (095) 727-91-19.
  
• E-mail: bushueva@yahoo.com
  
• Education: PhD since November 2001 in the field of molecular biology.
  
• Work at present: Head of molecular biology group at russian biomedical company "DNA-Technology".
  

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1. [Characterization of S1' subsite specificity of Thermoactinomyces vulgaris carboxypeptidase T by site-directed mutagenesis].
   [Article in Russian]
   Vopr Med Khim. 2002 Nov-Dec;48(6):577-85.
   Trachuk LA, Bushueva AM, Shevelev AB, Novgorodova SA, Akparov VKh, Chestukhina GG.

2. [Isolation of glutamylendopeptidase precursor from B. licheniformis and its processing in vitro].
   [Article in Russian]
   Vopr Med Khim 2001 Jan-Feb;47(1):111-22.
   Serkina AV, Bushueva AM, Chestukhina GG, Gumpert J, Hoischen C, Kujau M, Shevelev AB.

3. Expression of bacillar glutamyl endopeptidase genes in Bacillus subtilis by a new mobilizable single-replicon vector pLF.
   Plasmid 2000 May;43(3):190-9.
   Shevelev AB, Aleoshin VV, Trachuk LA, Granovsky AE, Kogan YN, Rumer LM, Serkina AV, Semenova EV, Bushueva AM, Livshits VA, Kostrov SV, Shcheglov AS, Novikova SI, Chestukhina GG.

4. Introduction of a mini-gene encoding a five-amino acid peptide confers erythromycin resistance on Bacillus subtilis and provides temporary erythromycin protection in Proteus mirabilis
   FEMS Microbiol Lett 2000 Jan 15;182(2):213-8.
   Novikova SI, Bushueva AM, Trachuk LA, Konstantinova GE, Serkina AV, Hoischen C, Gumpert J, Chestukhina GG, Mankin A, Shevelev AB.

5. Bacillus intermedius glutamyl endopeptidase. Molecular cloning and nucleotide sequence of the structural gene.
   J Protein Chem 1999 Jan;18(1):21-7.
   Rebrikov DV, Akimkina TV, Shevelev AB, Demidyuk IV, Bushueva AM, Kostrov SV, Chestukhina GG, Stepanov VM

6. Expression of the carboxypeptidase T gene from Thermoactinomyces vulgaris in stable protoplast type L-forms of Proteus mirabilis.
   FEMS Microbiol Lett. 1998 Feb 15;159(2):145-50.
   A.M. Bushueva, A.B. Shevelev, J. Gumpert, G.G. Chestukhina, A.V. Serkina, C. Hoischen, M.V. Matz, M.V. Kuryatova, V.M. Stepanov

7. A novel delta-endotoxin gene cryIM from Bacillus thuringiensis ssp. wuhanensis.
   FEBS Lett 1997 Mar 10;404(2-3):148-52.
   Shevelev AB, Kogan YaN, Bushueva AM, Voronina EJ, Rebrikov DV, Novikova SI, Chestukhina GG, Kuvshinov V, Pehu E, Stepanov VM

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• May 2003 - to present:
  Head of Molecular Biology group at russian biomedical company "DNA-Technology".
  
• 1993 - 2003
  Research Assistant, Protein Chemistry Lab. of Prof.Stepanov, Institute of Microbial Genetics GNIIgenetika.
  
• November - December, 1998
  An official participant of cooperativ research grant with Prof. J.Gumpert, Institute of Molecular Biotechnology, Jena, Germany.
  
• October 1997-December 1997
  An official participant of cooperativ research grant with Prof. J.Gumpert, Institute of Molecular Biotechnology, Jena, Germany.
  
• June-August 1997
  Summer student, Department of Membrain Research and Biophisics, Laboratory of Prof. D.Wallach, Weizmann Institute of Science, Rehovot, Israel.
  
• August-October, 1996
  An official participant of cooperative research project with Prof. J.Gumpert, Institute of Molecular Biotechnology, Jena, Germany.
  

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• 1999, 2000 and 2001
  President's PhD Scholarship for outstanding success in research and studies.
  
• 1996-to present
  Scholarship of International Soros Student Educational Programm for outstanding success in studies and research.
  
• 1993-1998
  All Russian Government Student Scholarships for outstanding success in studies.
  

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• Development of mutant variants of recombinant protease genes to adopt their expression and secretion for specific host, Escherichia coli and Bacillus subtilis (2000-at present), Protein Chemistry Laboratory, Institute of Microbial Genetics, Moscow, Russia
  Major techniques used: cloning of DNA fragments, PCR-based mutagenesis, gele electrophoresis, isolation of plasmid and genomic DNA from E.coli and B.subtilis, transformation of E.coli and B.subtilis cells, inoculation and maintaining of E.coli and B.subtilis strains expressing recombinant proteins, protein purification from inclusion bodies and from cultural liquid, SDS-PAAG electrophoresis, Western blot hybridization.
  
• PCR-based construction, expression in heterologous hosts and purification of mutant protein of carboxypeptidase T from Thermoactinomyces vulgaris (1999-at present), Protein Chemistry Laboratory, Institute of Microbial Genetics, Moscow, Russia
  Major techniques used: cloning of DNA fragments, sequencing, agarose gele-electroforesis, isolation of plasmid DNA, transformation of E.coli cells by calcium method and by electroporation, inoculation and maintaining of E.coli strains expressing recombinant protein, construction of cloning vectors for expression of mutant protein in Bacilli, protein purification from inclusion bodies of E.coli cells, SDS-PAAG electroforesis, Western blot hybridization, measurment of proteolytic activity of the protease on specific substrate.
  
• Expression of glutamylendopeptidase precursor from Bacillus licheniformis in L-forms of Proteus mirabilis (1998), Department of Procariotic Cell Biology of Institute of Molecular Biotechnology, Jena, Germany.
  Major techniques used: Western blot hybridization, inoculating and maintaining L-forms in liquid culture and on solid medium, microscopy, protein purification.
  
• Expression of glutamylendopeptidase from Bacillus licheniformis in L-forms of Proteus mirabilis (1997), Department of Procariotic Cell Biology of Institute of Molecular Biotechnology, Jena, Germany.
  Major techniques used: Western blot hybridization, inoculating and maintaining L-forms in liquid culture and on solid medium, microscopy, cloning of DNA, PCR-based exchange of gene parts, measurment of proteolytic activity of the protease on specific and non-specific substrate
  
• Study of genomic organization of murine gene NIK homologous to a man's gene involved in TNF induction and making a construct for knouck-out of the gene in mice (June - August 1996), Department of Membrane Research and Biophysics, Weizmann Institute of Science, Rehovot, Israel.
  Major techniques used: Southern blot hybridization, cloning and sequencing of cDNA and genomic DNA, PCR-based cloning, constructing of cDNA and genomic DNA libraries, making restriction and exon-intron map of genomic clones, making constructs for knock-out, agarose gele-electroforesis, isolation of phage DNA and plasmid DNA, transformation of E.coli cells by calcium method and by electroporation.
  
• Expression of carboxypeptidase T from Thermoactinomyces vulgaris in L-forms of Proteus mirabilis (1996), Department of Procariotic Cell Biology of Institute of Molecular Biotechnology, Jena, Germany.
  Major techniques used: Western blot hybridization, inoculating and maintaining L-forms in liquid culture and on solid medium, microscopy, cloning of DNA, measurment of proteolytic activity of the protease on specific and non-specific substrate
  
• Constructing of a new integrative cloning system for a wide range of Gram-positive hosts and system for cloning in Bacillus thuringiensis based on homolous recombination (1996-to present), Protein Chemistry Laboratory, Institute of Microbial Genetics, Moscow, Russia
  Major techniques used: cloning of DNA fragments, sequencing, conjugation of bacteria, Western blot hybridization, Southern blot hybridization of genomic DNA, isolation of total genomic DNA from Bacillus.
  
• Cloning and sequencing of delta-endotoxin genes of Bacillus thuringiensis (1994-1998), Protein Chemistry Laboratory, Institute of Microbial Genetics, Moscow, Russia.
  Major techniques used: constructing genomic libraries of bacteria, searching for new delta-endotoxin genes, Western blot hybridization, Southern blot hybridization, cloning of DNA fragments, sequencing, transformation of E.coli cells.
  

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A.M.Savilova is a participant of the following conferences:

• Oct. 2000
  Annual Conference of Federal science center GNIIgenetika: "Development of new expression sistems for bacterial proteinase genes in gram-positive and gram-negative hosts".
  
• Nov. 1999
  Conference for PhD students and young scientists, Institute of General Genetics of Russian Academy of Sciences and Department of Genetics and Selection of Moscow State University: "Study of carboxypeptidase T from T.vulgaris, the enzyme with unique double specificity".
  
• Oct. 1999
  Annual Conference of Federal science center GNIIgenetika: "Construction of high efficiency expression systems for production of bacterial proteinases and their application for practice use".
  
• Oct. 1998
  Anniversary Conference of Federal science center GNIIgenetika: "Construction of expression system for production of glutamylendopeptidase from Bacillus licheniformis in L-forms of Proteus mirabilis".
  
• Dec. 1997
  Conference of Institut fuer Moleculare Biotechnologie, Jena, Germany: "Results of the investigations concerning the expression of and glutamate-specific protease from Bacillus licheniformis in L-forms of Proteus mirabilis".
  
• Oct. 1997
  V Scientific Conference of Federal scientific center GNIIgenetika: "Expression of carboxypeptidase T gene from Òhermoactinomyces vulgaris in L-form cells of Proteus mirabilis".
  
• Aug. 1997
  Conference of Weizmann Institute of Science, Israel, Rehovot, report: "Study of genomic organisation of mouse gene NIK involved in TNF signaling to make its knockout in mice".
  
• April 21-23 1997
  IV State Simposium "Chemistry of Proteolytic enzymes", Moscow, Institute of Bioorganic Chemistry, "Expression of carboxypeptidase T gene from Òhermoactinomyces vulgaris in L-form cells of Proteus mirabilis".
  
• Oct. 1996
  Conference of Institute fuer Moleculare Biotechnologie, Jena, Germany, "Results of the investigations concerning the expression of carboxypeptidase T and glutamate-specific protease in L-forms of Proteus mirabilis".
  
• Apr. 1996
  Annual conference of Federal science center GNIIgenetika: "Construction of integrative system for cloning in wide range of Gram-positive hosts on the basis of transposon-like element of Bacillus thuringiensis wuhanensis".
  
• Feb. 1996
  International conference "Plant resistance to biotic and abiotic stresses" Helsinki univercity, Helsinki, Finland, "New approaches to identification and cloning of new delta-endotoxin genes from Bacillus thuringiensis".
  
• Jan. 1995
  Annual conference of Federal scientific center GNIIgenetika, "Study of genomic organization of delta-endotoxins from Bacillus thuringiensis".
  
• Jan. 1995
  Annual conference of Federal scientific center GNIIgenetika: "Construction of a new single-replicone mobilized shuttle-vector for cloning in a wide range of Gram-positive hosts".
  
• May-June 1995
  New guide-lines in biotechnology, Pushchino: "Study of structural-functional organization of delta-endotoxins of Bacillus thuringiensis by a method of molecular hybrids".
  
• May 1994
  New guide-lines in biotechnology, Pushchino, "Study of delta-endotoxin organization of Bacillus thuringiensis by protein engineering methods".
  
• Nov. 1994
  "Study of structural-functional organization of crystal proteins-entomotoxins", conference on program "New methods of bioengineering. Protein engineering", Moscow, Institute of Bioorganic Chemistry.
  
• 6-10 Dec. 1994
  International conference "Plant resistance to biotic and abiotic stresses" Helsinki univercity, Helsinki, Finland, "Genomic organization of delta-endotoxin genes of Bacillus thuringiensis".
  
• Nov. 1993
  "Study of the structural-functional organization of entomocidal proteins-entomotoxins", annual conference on a state scientific program "New approaches in bioengineering. Protein engineering", Moscow, Institute of Bioorganic Chemistry.
  

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