Cloning of human insulin-like growth factor gene in plant expression vectors

Abstract

Transgenic plants are suitable bioreactors for the production of recombinant proteins at high levels and qualities. Cloning of a gene encoding the target protein in proper expression vectors is one of the most important steps for successful integration and expression of target protein in plants. The purpose of this study was to clone insulin-like growth factor -1 (IGF1) cDNA in plant expression vectors. To this end, we amplified amplify IGF1 cDNA using a pair of specific primer followed by its cloning in pMCS5 basic vector. After sequencing, because of several advantages of cereals for production of biopharmaceuticals, such as high cultivation area and biomass, possibility of grain storage and usage as food and feed, we constructed suitable vectors for IGF1 integration and expression in cereals. In addition, as transgenic chloroplasts contain higher potential to produce elevated levels of functional recombinant proteins, the IGF1 cDNA was recloned in two chloroplast-specific vectors. In one of these vectors, selectable marker gene is flanked by two direct repeats of LoxP sequences, providing the marker gene excision after production of transgenic plants via Cre/Lox system.

Keywords

• : insulin-like growth factor -1
• cloning
• gene transfer
• recombinant proteins
• expression vectors