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Cloning and expression of human bone morphogenetic protein 2 gene in Leishmania tarentolae - دانلود رایگان



دانلود رایگان Cloning and expression of human bone morphogenetic protein-2 gene in Leishmania tarentolae

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Cloning and expression of human bone morphogenetic protein-2 gene in Leishmania tarentolaeنوع فایل:PDF
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سال انتشار:1395
چکیده
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 is
simple and robust, but inept to make disulfide bond; hence, heterologous proteins are produced in
the form of inclusion bodies, entailing in vitro refolding strategies. On the other hand, yeast, which
is a lower eukaryote, is able to perform glycosylation of the recombinant proteins. However, its
glycosylation pattern is different from human beings. Ultimately, Leishmania tarentolae, a parasite
protozoan of the lizard, unifying both the characteristics of prokaryotes and eukaryotes, has been
introduced for the expression of glycoproteins. It is capable of making disulfide bonds, expressing
correctly folded and biologically active recombinant proteins. In the current study, we successfully
cloned and expressed proBMP-2 gene in L. tarentolae. Integration of the gene was confirmed by
PCR; 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 kDa
band was obtained, equivalent to monomeric form of BMP-2.
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