title: Chemical synthesis and improved  expression of recombinant human granulocyte colony-stimulating factor cDNA.
abstract: Recently,  granulocyte colony-stimulating factor (G-CSF) has been recognized as a useful  molecule for the treatment of a wide range of complex ailments, such as cancer,  AIDS, H1N1 influenza, cardiac and neurological diseases. The vast therapeutic  potential of G-CSF has induced scientists to develop biotechnological  approaches for the synthesis of this pharmacologically active agent. We used a  synthetic G-CSF cDNA molecule to produce the target protein by a simple cloning  protocol. We constructed the synthetic cDNA using a DNA synthesizer with the  aim to increase its expression level by specific sequence modifications at the  5' end of the G-CSF-coding region, decreasing the GC content without altering  the predicted amino acid sequences. The identity of the resulting protein was  confirmed by a highly specific enzyme-linked immunosorbent assay. In  conclusion, a synthetic G-CSF cDNA in combination with the recombinant DNA  protocol offers a rapid and reliable strategy for synthesizing the target  protein. However, commercial utilization of this methodology will require  rigorous validation and quality control.
authors: King Abdullah Institute for Nanotechnology, King Saud University, Riyadh, Saudi Arabia. dr.salman@alrokayan.com
authors: S
authors: S
authors: Alrokayan
mesh_terms: D003001:Cloning, Molecular / Q000379:methods; D018076:DNA, Complementary / Q000138:chemical synthesis* / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D016179:Granulocyte Colony-Stimulating Factor / Q000096:biosynthesis* / Q000235:genetics; D006801:Humans; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics
chemical_list: D018076:DNA, Complementary; D011994:Recombinant Proteins; D016179:Granulocyte Colony-Stimulating Factor