expression and purification of human interferon gamma using a plant viral vector

نویسندگان

sayed mohsen nassaj hosseini

masoud shams-bakhsh

ali-hatef salamanian

shyi-dong yeh

چکیده

a plant viral vector engineered from an in vivo infectious clone of zucchini yellow mosaic virus(zymv) was used to express the human interferon-gamma (inf-γ) in planta. the inf-γ gene was in frame inserted between the p1 and hc-pro orfs of the zymv vector. the infectious activity of the vector was approved by rubbing the plasmid on chenopodium quino a and observing local lesions. individual lesions were mechanically transferred to the systemic host plant zucchini squash at the stage of cotyledonary leaf. the stability of inf-γ expression was assessed by successive passages of recombinant viruses from infected plant and throughout the period of 35 days after inoculating in a single plant. then, the leaf tissues ofinoculated plant were analyzed for the presence of transgene by rt-pcr and western blot analysis. the recombinant protein was purified using affinity chromatography method. the results showed approximately 1–1.2 mg inf-γ per 100 g tissues were purified from leaves two weeks post inoculation. also, the vector was remarkably stable in squash after six serial passages and 35 days. the procedure provides a convenient and fast method for production of large quantities of pure inf-γin planta. the system also has a potential for production of other proteins of interest in cucurbits to use as immunogen to produce antiserum or use for other purposes.

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

Expression and Purification of Human Interferon Gamma Using a Plant Viral Vector

A plant viral vector engineered from an in vivo infectious clone of zucchini yellow mosaic virus(ZYMV) was used to express the human interferon-gamma (INF-γ) in planta. The INF-γ gene was in frame inserted between the P1 and HC-Pro ORFs of the ZYMV vector. The infectious activity of the vector was approved by rubbing the plasmid on Chenopodium quino a and observing local lesions. Individual les...

متن کامل

Expression and purification of human interferon gamma using a plant viral vector

A plant viral vector engineered from an in vivo infectious clone of zucchini yellow mosaic virus (ZYMV) was used to express the human interferon-gamma (INF-γ) in planta. The INF-γ gene was in frame inserted between the P1 and HC-Pro ORFs of the ZYMV vector. The infectious activity of the vector was approved by rubbing the plasmid on Chenopodium quinoa and observing local lesions. Individual les...

متن کامل

Inducible Expression of Human Gamma Interferon

Background:The premature termination of high producer clones, which will be killed due to cell proliferation and proteins production antagonism, is one of the basic drawback in recombinant proteins technology. Furtheremore, it is supposed some toxic proteins like interferon which we intended to clone and express, inhibit host cells’ proliferation. So, it is necessary to tightly control IFN-γ pr...

متن کامل

inducible expression of human gamma interferon

background:the premature termination of high producer clones, which will be killed due to cell proliferation and proteins production antagonism, is one of the basic drawback in recombinant proteins technology. furtheremore, it is supposed some toxic proteins like interferon which we intended to clone and express, inhibit host cells’ proliferation. so, it is necessary to tightly control ifn-γ pr...

متن کامل

Cloning and Expression of Human Gamma-Interferon cDNA in E. coli

Prior to the production of human gamma interferon using recombinant DNA technology, it had been producedmainly upon mitogenic induction of lymphocytes in very low amounts, which evidently hamperedits characterization and its medical applications. The recombinant gamma interferons produced in largerquantities in prokaryotic systems retain their biological activities, and can be...

متن کامل

A Simplified Process for Purification and Refolding of Recombinant Human Interferon-α2b

Background: Interferon α-2b is a vital biotherapeutic produced through the recombinant DNA technology in E. coli. The recombinant IFN-α2b normally appears as intercellular IBs, which requires intensive refolding and purification steps. Method: Purification of IFN-α2b from solubilized IB was performed using two-phase extraction. To optimize refolding conditions, the effects of pH and different a...

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:
progress in biological sciences

ناشر: university of tehran

ISSN 1016-1058

دوره 2

شماره 2 2013

کلمات کلیدی

میزبانی شده توسط پلتفرم ابری doprax.com

copyright © 2015-2023