Proteins in a DNA world: expression systems for their study.

Every day, new proteins are discovered and the need to understand its function arises. Human proteins have a special interest, because to know its role in the cell may lead to the design of a cure for a disease. In order to obtain such information, we need enough protein with a high degree of purity, and in the case of the human proteins, it is almost impossible to achieve this by working on human tissues. For that reason, the use of expression systems is needed. Bacteria, yeast, animals and plants have been genetically modified to produce proteins from different species. Even "cell-free" systems have been developed for that purpose. Here, we briefly review the options with their advantages and drawback, and the purification systems and analysis that can be done to gain understanding on the function and structure of the protein of interest.

[1]  Y. Schneider,et al.  Insect cell culture for industrial production of recombinant proteins , 2003, Applied Microbiology and Biotechnology.

[2]  A. Brake.  [34] α-Factor leader-directed secretion of heterologous proteins from yeast , 1990 .

[3]  I. Chung,et al.  Optimization of transfection conditions for expression of green fluorescent protein in Drosophila melanogaster S2 cells , 1999 .

[4]  K. Ohashi,et al.  Expression of two subtypes of human IFN-alpha in transgenic potato plants. , 2001, Journal of interferon & cytokine research : the official journal of the International Society for Interferon and Cytokine Research.

[5]  K. Gevaert,et al.  Protein identification methods in proteomics , 2000, Electrophoresis.

[6]  H. Sang,et al.  Transgenic: birds by DNA microinjection , 1994 .

[7]  R. B. Trimble,et al.  Overview of N- and O-linked oligosaccharide structures found in various yeast species. , 1999, Biochimica et biophysica acta.

[8]  M. J. Chalmers,et al.  Advances in mass spectrometry for proteome analysis. , 2000, Current opinion in biotechnology.

[9]  A. Spirin,et al.  A continuous cell-free translation system capable of producing polypeptides in high yield. , 1988, Science.

[10]  Julie A. Morris,et al.  Expression of exogenous protein in the egg white of transgenic chickens , 2002, Nature Biotechnology.

[11]  K. Ohashi,et al.  Expression of Two Subtypes of Human IFN-α in Transgenic Potato Plants , 2001 .

[12]  J. Kane,et al.  Effects of a minor isoleucyl tRNA on heterologous protein translation in Escherichia coli , 1995, Journal of bacteriology.

[13]  C. Kurland.  Codon bias and gene expression , 1991, FEBS letters.

[14]  A. Brake.  Alpha-factor leader-directed secretion of heterologous proteins from yeast. , 1990, Methods in enzymology.

[15]  A. Panda,et al.  Bioprocessing of therapeutic proteins from the inclusion bodies of Escherichia coli. , 2003, Advances in biochemical engineering/biotechnology.

[16]  T. Park,et al.  Effect of silkworm hemolymph on N‐linked glycosylation in two Trichoplusia ni insect cell lines , 2003, Biotechnology and bioengineering.

[17]  Ajay Singh,et al.  Developments in the use of Bacillus species for industrial production. , 2004, Canadian journal of microbiology.

[18]  T. Sawasaki,et al.  Construction of an efficient expression vector for coupled transcription/translation in a wheat germ cell-free system. , 2000, Nucleic acids symposium series.

[19]  E. Laskowska,et al.  Transcription of the ibpB Heat-Shock Gene Is under Control of ς32- and ς54-Promoters, a Third Regulon of Heat-Shock Response , 2001 .

[20]  E. Pauwels,et al.  Large scale production of recombinant human lactoferrin in the milk of transgenic cows , 2002, Nature Biotechnology.

[21]  S J Ward,et al.  Impact of genomics in drug discovery. , 2001, BioTechniques.

[22]  Harish Padh,et al.  Expression systems for production of heterologous proteins , 2001 .

[23]  H. A. Wood,et al.  Comparative recombinant protein production of eight insect cell lines , 1993, In Vitro Cellular & Developmental Biology - Animal.

[24]  A. Helenius,et al.  Intracellular functions of N-linked glycans. , 2001, Science.

[25]  S. Chong,et al.  Fusions to self-splicing inteins for protein purification. , 2000, Methods in enzymology.

[26]  J-M Betton,et al.  Rapid translation system (RTS): a promising alternative for recombinant protein production. , 2003, Current protein & peptide science.

[27]  W. Vos.  Gene expression systems for lactic acid bacteria. , 1999 .

[28]  H. Robertson,et al.  Factors affecting transposition of the Himar1 mariner transposon in vitro. , 1998, Genetics.

[29]  R. Dwek,et al.  Glycosylation and the immune system. , 2001, Science.

[30]  K. Terpe.  Overview of tag protein fusions: from molecular and biochemical fundamentals to commercial systems , 2002, Applied Microbiology and Biotechnology.

[31]  K. Simons,et al.  Protein expression in Drosophila Schneider cells. , 2000, Analytical biochemistry.

[32]  J. Carrington,et al.  A viral cleavage site cassette: identification of amino acid sequences required for tobacco etch virus polyprotein processing. , 1988, Proceedings of the National Academy of Sciences of the United States of America.

[33]  E. Laskowska,et al.  Transcription of the ibpB heat-shock gene is under control of sigma(32)- and sigma(54)-promoters, a third regulon of heat-shock response. , 2001, Biochemical and biophysical research communications.

[34]  R. Haun,et al.  Ligation-independent cloning of glutathione S-transferase fusion genes for expression in Escherichia coli. , 1992, Gene.

[35]  R. Ivarie,et al.  Consistent production of transgenic chickens using replication-deficient retroviral vectors and high-throughput screening procedures. , 2002, Poultry science.

[36]  K. Nagai,et al.  Generation of β-globin by sequence-specific proteolysis of a hybrid protein produced in Escherichia coli , 1984, Nature.

[37]  R. Ivarie.  Avian transgenesis: progress towards the promise. , 2003, Trends in biotechnology.

[38]  J. Cregg,et al.  Applications of yeast in biotechnology: protein production and genetic analysis. , 1999, Current opinion in biotechnology.

[39]  C. Kurland,et al.  Co-variation of tRNA abundance and codon usage in Escherichia coli at different growth rates. , 1996, Journal of molecular biology.

[40]  International Human Genome Sequencing Consortium.  Initial sequencing and analysis of the human genome , 2001, Nature.

[41]  D. J. Bell.  Physiology and Biochemistry of the Domestic Fowl , 1971 .

[42]  C. Scorer,et al.  Foreign gene expression in yeast: a review , 1992, Yeast.

[43]  P. Stephens,et al.  High-level periplasmic expression in Escherichia coli using a eukaryotic signal peptide: importance of codon usage at the 5' end of the coding sequence. , 2000, Protein expression and purification.

[44]  H. Koprowski,et al.  Expression in plants and immunogenicity of plant virus-based experimental rabies vaccine. , 2002, Vaccine.

[45]  M. Huddleston,et al.  Mistranslational errors associated with the rare arginine codon CGG in Escherichia coli. , 2003, Protein expression and purification.

[46]  B. Coddeville,et al.  Comparative analysis of the site-specific N-glycosylation of human lactoferrin produced in maize and tobacco plants. , 2003, European journal of biochemistry.

[47]  C. Fuqua,et al.  Broad-host-range expression vectors that carry the L-arabinose-inducible Escherichia coli araBAD promoter and the araC regulator. , 1999, Gene.

[48]  W. D. de Vos,et al.  Gene expression systems for lactic acid bacteria. , 1999, Current opinion in microbiology.

[49]  J. Cregg,et al.  Production of recombinant proteins in fermenter cultures of the yeast Pichia pastoris. , 2002, Current opinion in biotechnology.

[50]  R. Fletterick,et al.  Characterization of the catalytic residues of the tobacco etch virus 49-kDa proteinase. , 1989, Virology.

[51]  J. Carrington,et al.  Biochemical and mutational analysis of a plant virus polyprotein cleavage site. , 1988, The EMBO journal.

[52]  D. McRee.  Practical Protein Crystallography , 1993 .

[53]  K. Nagai,et al.  Generation of beta-globin by sequence-specific proteolysis of a hybrid protein produced in Escherichia coli. , 1984, Nature.

[54]  N. Hilschmann,et al.  Thrombin specificity. Selective cleavage of antibody light chains at the joints of variable with joining regions and joining with constant regions. , 1985, European journal of biochemistry.