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Title of Journal: Mol Biotechnol

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Abbravation: Molecular Biotechnology

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Springer US

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DOI

10.1007/s00277-015-2339-6

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ISSN

1559-0305

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Codon Optimization of the “Bos Taurus Chymosin” Ge

Authors: Jose Alberto EspinozaMolina Carlos Horacio AcostaMuñiz D R Sepulveda Paul Baruk ZamudioFlores Claudio RiosVelasco
Publish Date: 2016/07/09
Volume: 58, Issue: 10, Pages: 657-664
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Abstract

Codon optimization of the Bos taurus Chymosin gene CYM for its expression in Pichia pastoris was performed in this study A synthetic CYM gene was designed in silico by replacing codons rarely used by P pastoris with equivalent nucleotide combinations that codify for the same amino acid but that are more frequently encountered in the genome of P pastoris A total of 332 nucleotides were modified to optimize 289 codons The synthetic CYM gene was cloned into the expression vector pPICZαA and transformed into P pastoris The transformed strains were grown in artificial media supplemented with glycerol as a carbon source to increase biomass and then cultured in a similar medium replacing glycerol with methanol as a carbon source to initiate gene induction Raw extracts of the growth media exhibited milkclotting activity of 14611 SU/mL Produced recombinant chymosin showed coagulant activity from 25 to 50 °C and within a pH range of 5–69 having optimum activity at 35–40 °C and pH 50 These results show that codon optimization is a viable strategy to improve CYM gene expression levels in P pastoris for the production of recombinant chymosin

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  1. Production and Characterization of a Single-Chain Fv Antibody–Alkaline Phosphatase Fusion Protein Specific for Clenbuterol
  2. Improvement of Cephalosporin C Production by Recombinant DNA Integration in Acremonium chrysogenum
  3. Heterologous Overexpression, Purification and Characterisation of an Alcohol Dehydrogenase (ADH2) from Halobacterium sp. NRC-1
  4. Enhanced Extracellular Production of Heterologous Proteins in Bacillus subtilis by Deleting the C-terminal Region of the SecA Secretory Machinery
  5. Enhanced Production of Human Recombinant Proteins from CHO cells Grown to High Densities in Macroporous Microcarriers
  6. Molecular Cloning and Characterization of a Malic Enzyme Gene from the Oleaginous Yeast Lipomyces starkeyi
  7. Hydrophobic Substitution of Surface Residues Affects Lipase Stability in Organic Solvents
  8. Identification of Suitable Endogenous Normalizers for qRT-PCR Analysis of Plasma microRNA Expression in Essential Hypertension
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  10. Heterologous Expression and Characterization of an Alcohol Dehydrogenase from the Archeon Thermoplasma acidophilum
  11. Plasma Antioxidants and Human Aging: A Study on Healthy Elderly Tunisian Population
  12. Importance of the residue Asp 290 on chain length selectivity and catalytic efficiency of recombinant Staphylococcus simulans lipase expressed in E. coli
  13. Fahd Al-Mulla (ed). Formalin-Fixed Paraffin-Embedded Tissue. Methods and Protocols. Methods in Molecular Biology 724. The Humana Press, ISBN 978-1-61779-054-6
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  15. End Joining-Mediated Gene Expression in Mammalian Cells Using PCR-Amplified DNA Constructs that Contain Terminator in Front of Promoter
  16. Identification of Functional Regions in the Rhodospirillum rubrum l -Asparaginase by Site-Directed Mutagenesis
  17. Expression of an Acid Urease with Urethanase Activity in E. coli and Analysis of Urease Gene
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