Cloning and expression of human bone morphogenetic protein-2 gene inLeishmania tarentolae

Publish Year: 1395
نوع سند: مقاله کنفرانسی
زبان: English
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شناسه ملی سند علمی:

ICESCON03_391

تاریخ نمایه سازی: 16 شهریور 1395

Abstract:

Several expression hosts have been employed for the production of bone morphogenetic protein-2 (BMP- 2), inducing de novo bone formation and is widely used in clinical applications. However,every expression system has its merits and demerits. For example, although Escherichia coli issimple and robust, but inept to make disulfide bond; hence, heterologous proteins are produced inthe form of inclusion bodies, entailing in vitro refolding strategies. On the other hand, yeast, whichis a lower eukaryote, is able to perform glycosylation of the recombinant proteins. However, itsglycosylation pattern is different from human beings. Ultimately, Leishmania tarentolae, a parasiteprotozoan of the lizard, unifying both the characteristics of prokaryotes and eukaryotes, has beenintroduced for the expression of glycoproteins. It is capable of making disulfide bonds, expressingcorrectly folded and biologically active recombinant proteins. In the current study, we successfullycloned and expressed proBMP-2 gene in L. tarentolae. Integration of the gene was confirmed byPCR; whereas, transcription of the coding gene was verified by back translating the target mRNA.Besides, the expression of BMP-2 was assessed by SDS-PAGE and Western blotting. A 12 kDaband was obtained, equivalent to monomeric form of BMP-2.

Keywords:

Bone morphogenetic protein 2 , Morphogenesis , Growth differentiation factor 2 , Osteogenesis , Spinal cord injuries , Glycosylation

Authors

Majid Rahmati

Department of Medical Biotechnology, School of Advanced Technologies in Medicine, Tehran

Amjad Hayat Khan

Department of Medical Biotechnology, School of Advanced Technologies in Medicine, InternationalCampus, Tehran University of Medical Sciences, Tehran, Iran

Shabnam Razavi

Department of Medical Biotechnology, School of Advanced Technologies in Medicine, Tehran

Mohammad Reza Khorramizadeh

Department of Medical Biotechnology, School of Advanced Technologies in Medicine, Tehran