a comparison of bone morphogenetic protein-7 mutant expression in prokaryotic and eukaryotic hosts

نویسندگان

leila nematollahi

fereidoun mahboudi

mohammad azizi

farzaneh barkhordari

ahmad adeli

چکیده

objective: bone morphogenetic protein-7 (bmp-7) is a multifunctional growth factor predominantly recognized for its osteoinductive properties. due to the high cost of this protein, the availability of bmp-7 for treatment is limited. the heterologous production of recombinant hbmp-7 has been performed in a number of expression systems. in this study a novel form of bmp-7 was expressed in eukaryotic and prokaryotic hosts. methods: for expression in the prokaryotic system, the novel protein was secreted to the periplasmic space of escherichia coli using a pelb signal sequence followed by single-step purification by ni2+-charged column chromatography. in the mammalian cell expression system, we transferred a full-length cdna encoding precursor of the novel protein to cho cells then selected stable clones by using the appropriate antibiotic concentration. expressions in both systems were confirmed by western blot analysis. results: the novel recombinant protein was produced as a 36-38 kda dimer in the cho cell line and a 16 kda monomer in the escherichia coli system. quantitative analysis according to elisa showed that the expression levels of the mutant protein in the eukaryotic and prokaryotic expression systems were 40 ng/ml and 135 ng/ml of the culture media, respectively. conclusion: in this study, the expression level in escherichia coli was at least three times more than observed in the cho cells. however, further optimization is required to obtain a dimer protein in escherichia coli. the results show that periplasmic expression may be suitable for the production of complex proteins such as bmps.

برای دانلود باید عضویت طلایی داشته باشید

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

periplasmic expression of a novel human bone morphogenetic protein-7 mutant in escherichia coli

background: bone morphogenetic proteins (bmps) belong to the transforming growth factor-β (tgf-β) superfamily, and play an important role in bone metabolism. recombinant forms of bmp-2 and bmp-7 are the only bmps used clinically. in this study the mature part of human bone morphogenetic protein-7 (bmp-7) was engineered through substitution of the bmp-7 n-terminal sequence by heparin-binding sit...

متن کامل

Periplasmic Expression of a Novel Human Bone Morphogenetic Protein-7 Mutant in Escherichia coli

BACKGROUND Bone Morphogenetic Proteins (BMPs) belong to the transforming growth factor-β (TGF-β) superfamily, and play an important role in bone metabolism. Recombinant forms of BMP-2 and BMP-7 are the only BMPs used clinically. In this study the mature part of human bone morphogenetic protein-7 (BMP-7) was engineered through substitution of the BMP-7 N-terminal sequence by heparin-binding site...

متن کامل

changes in the expression of bone morphogenetic protein 7 and tamm– horsfall protein in the early stages of diabetic nephropathy

background bone morphogenetic protein 7 (bmp7) has been suggested to play a protective role against kidney injury in chronic kidney disease. objectives to identify the critical molecular regulators in the early stage of diabetic nephropathy, we studied the expression of bmp7 and 2 important kidney-specific markers, podocin and tamm–horsfall protein (thp). materials and methods a diabetic nephro...

متن کامل

Bone morphogenetic protein-7 expression in human pyelonephritis.

Bone morphogenetic protein 7 (BMP- 7) is a member of the transforming growth factor (TGF) beta superfamily and is involved in regeneration, repair, and development of specific tissues, for example kidney and skeleton. The experimental studies have shown its protective role against fibrotic processes. Tubulointerstitial changes are present in the pyelonephritic kidney which progresses to fibrosi...

متن کامل

assessment of deep word knowledge in elementary and advanced iranian efl learners: a comparison of selective and productive wat tasks

testing plays a vital role in any language teaching program. it allows teachers and stakeholders, including program administrators, parents, admissions officers and prospective employers to be assured that the learners are progressing according to an accepted standard (douglas, 2010). the problems currently facing language testers have both practical and theoretical implications but the first i...

Over Expression of Influenza Virus M2 Protein in Prokaryotic System

Background and Aims: Influenza A virus of Orthomyxoviridae family is able to create pandemic influenza. Vaccination is the most effective way to prevent influenza virus infection. Matrix protein 2 (M2) is a homotetramer ion channel with 97 amino acids length and highly conserved among influenza viruses and is considered for development of a universal influenza vaccine. Materials and Methods: We...

متن کامل

منابع من

با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید


عنوان ژورنال:
modares journal of medical sciences: pathobiology

ناشر: tarbiat modares university

ISSN 1562-9554

دوره 15

شماره 3 2012

میزبانی شده توسط پلتفرم ابری doprax.com

copyright © 2015-2023