Isolation and Cloning of Human Insulin - Like Growth Factor 1 ( IGF 1 ) Cdna in Chloroplast Transformation Vectors

Abstract

Diabetes is one of the most common chronic troubles in the world that occurs when the body fails to process sugar correctly, largely because of inadequate insulin production. At present, insulin injection is the most prominent method for adequate insulin replacement. Unfortunately, pain and discomfort condition of the self-injection system leads to inaccurate injection of insulin. To solve such problems, oral administration is suggested. However this method increases conformity of therapy, it typically requires 5-20 fold higher insulin doses. From other side, incidence of diabetes is rapidly increasing worldwide. Therefore, more economical methods should be considered to produce sufficient amounts of insulin. Plant systems are one of the most efficient methods to produce many proteins at industrial levels. Therefore, in this project, we aimed to produce insulin in plant systems. We first isolated the cDNA of IGF1 from cultured human Hell cells using a pair of specific primers. Isolated cDNA is then cloned in basic vectors, followed by sequencing. Confirmed cDNA sequences are then cloned in different plant transformation vectors suitable for nuclear transformation via Agrobacterium or Biolistic systems, as well for plastid transformation. As edible plants with a high biomass can be suitable systems for production of insulin at a higher level with minimized downstream protein purification, construction of lettuce plastid-specific transformation vector is being considered. Also, because highly efficient plant regeneration systems are of pivotal importance for recovering plastid-tranformed plants, we have developed a highly responsive leaf-based plant regeneration system for lettuce. Keywords—Molecular cloning, Insulin, Diabetes, Recombinant protein

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Cite this paper

@inproceedings{NojavanIsolationAC, title={Isolation and Cloning of Human Insulin - Like Growth Factor 1 ( IGF 1 ) Cdna in Chloroplast Transformation Vectors}, author={S . Nojavan and Mohammad Ahmadabadi and Mehrdad Pazhouhandeh} }