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Iranian Journal of Basic Medical Sciences، جلد ۱۸، شماره ۸، صفحات ۷۸۰-۷۸۷
عنوان فارسی
چکیده فارسی مقاله
کلیدواژه‌های فارسی مقاله
عنوان انگلیسی Construction of expressing vectors including melanoma differentiation-associated gene-7 (mda-7) fused with the RGD sequences for better tumor targeting
چکیده انگلیسی مقاله Objective(s): Up to now, many researches have been performed to improve the antitumoral effect of melanoma differentiation-associated gene-7 (mda-7) protein. The purpose of our research was to construct 3 expression vectors producing mda-7 in fusion with RGD (Arginine-Glycine-Aspartic acid) peptide and evaluate their expression. Materials and Methods: mda-7 gene with two different RGD sequences was amplified by PCR then was cloned by TA–cloning system. The colonies including these genes were selected by blue–white screening, colony PCR, and sequencing, respectively. Afterward, the genes were sub-cloned into the expression vector following confirmation by colony PCR and sequencing. In addition, these constructs were transfected into 293 and Huh-7 cells for further expression analysis. The mda-7 gene expression was evaluated by RT-PCR and IF (immunofluorescence assay). DNA laddering test and trypan blue exclusion assays were performed to screen cytotoxicity of prepared plasmids. Results: Three different mda-7 genes with terminal RGD peptide were cloned correctly into the expression vectors and their expression was confirmed to be suitable by RT-PCR and IF assay. It was shown that expressions were limited to those transfected, GFP shining cells. No significant cytotoxicity was observed by simple assays in all plasmid treated cells. In expressing cells, all forms of mda-7 protein were localized mainly around ER prenuclear compartment while GFP protein was distributed evenly among them. Conclusion: Theoretically RGD tagged mda-7 would be able to induce apoptosis with more specificity and stronger than the standard one, therefore, these new constructs may have the potential for further researches.
کلیدواژه‌های انگلیسی مقاله Mda-7, RGD peptide, Tumor targeting
نویسندگان مقاله محبوبه خداداد | mahboobeh khodadad
gastroenterohepatology research center, shiraz university of medical sciences, shiraz, iran

سازمان اصلی تایید شده: دانشگاه علوم پزشکی شیراز (Shiraz university of medical sciences)

سید یونس حسینی | seyed younes hosseini
gastroenterohepatology research center, shiraz university of medical sciences, shiraz, iran

سازمان اصلی تایید شده: دانشگاه علوم پزشکی شیراز (Shiraz university of medical sciences)

فاطمه شناور | fatemeh shenavar
gastroenterohepatology research center, shiraz university of medical sciences, shiraz, iran

سازمان اصلی تایید شده: دانشگاه علوم پزشکی شیراز (Shiraz university of medical sciences)

نصراله عرفانی | nasrollah erfani
cancer immunology research group, shiraz institute for cancer research, school of medicine, shiraz university of medical sciences, shiraz, iran

سازمان اصلی تایید شده: دانشگاه علوم پزشکی شیراز (Shiraz university of medical sciences)

سمانه بینا | samaneh bina
gastroenterohepatology research center, shiraz university of medical sciences, shiraz, iran|sciences institute of biochemistry and biophysics, university of tehran, tehran, iran

سازمان اصلی تایید شده: مرکز تحقیقات بیوشیمی و بیوفیزیک (Institute of biochemistry and biophysics)

شاهین احمدیان | shahin ahmadian
sciences institute of biochemistry and biophysics, university of tehran, tehran, iran

سازمان اصلی تایید شده: مرکز تحقیقات بیوشیمی و بیوفیزیک (Institute of biochemistry and biophysics)

محمدرضا فتاحی | mohammad reza fattahi
gastroenterohepatology research center, shiraz university of medical sciences, shiraz, iran

سازمان اصلی تایید شده: دانشگاه علوم پزشکی شیراز (Shiraz university of medical sciences)

رضا حاج حسینی | reza hajhosseini
department of biochemistry, payame noor university, tehran shargh branch, tehran, iran

سازمان اصلی تایید شده: دانشگاه پیام نور تهران (Payame noor university)

نشانی اینترنتی http://ijbms.mums.ac.ir/article_4729.html
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