چاپ صفحه |  صفحه نخست سامانه |  چکیده مقاله |  نویسندگان |  دانلود مقاله | دانشگاه علوم پزشکی تبریز |
| نویسنده ثبت کننده مقاله | صفر فرج نیا |
| مرحله جاری مقاله | تایید نهایی |
| دانشکده/مرکز مربوطه | مرکز تحقیقات کاربردی دارویی |
| کد مقاله | 62322 |
| عنوان فارسی مقاله | بیالن محلول TNF-a انسانی با واسطه چاپرون DsbC |
| عنوان لاتین مقاله | DsbC chaperone mediated soluble expression of human TNF-a ٍE. coli |
| ناشر | 5 |
| آیا مقاله از طرح تحقیقاتی و یا منتورشیپ استخراج شده است؟ | بلی |
| عنوان نشریه (خارج از لیست فوق) | |
| نوع مقاله | Original Article |
| نحوه ایندکس شدن مقاله | ایندکس شده سطح یک – ISI - Web of Science |
| آدرس لینک مقاله/ همایش در شبکه اینترنت | https://www.minervamedica.it/en/journals/minerva-biotecnologica/article.php?cod=R04Y2018N01A0014 |
| Background: TNF-a is a pleiotropic cytokine that plays an important role in both physiological and pathological conditions. An increasing demand of large amount of bioactive TNF for use in clinic and drug development strategies make this cytokine as one of the most important biotechnologically produced pharmaceuticals. In this study, soluble expression of TNF-a in cytoplasm of E. coli was attempted using Dsbc chaperon and the effects of environmental condition including inducer concentration and incubation temperature was evaluated. Method: The gene encoding TNF-a was isolated by PCR reaction from cDNA that generated from total RNA extracted from PBMC and cloned into the pET-22b expression vector. Two type of E. coli strains, IPTG concentrations and growth temperatures as well as different type of media were examined with respect to the amount of recombinant protein produced in soluble form. Cytotoxicity assay on L929 cell line was performed for bioactivity evaluation. Results: Maximum amount of soluble TNF-a was attained by induction with 0.2 mM IPTG at 25 °C in the LB media. MTT assay with recombinant protein on L929 cells showed TNF-a expressed in active form with EC50 of 112pg/ml. Conclusion: This study report Shuffle strain markedly increases soluble expression of TNF-α .in optimized condition. |
| نویسنده | نفر چندم مقاله |
|---|---|
| حسن داریوش نژاد | اول |
| صفر فرج نیا | دوم |
| نصرت اله ضرغامی | سوم |
| شیوا عهدی خسروشاهی | چهارم |
| لیلا رهبرنیا | پنجم |
| نام فایل | تاریخ درج فایل | اندازه فایل | دانلود |
|---|---|---|---|
| Dariushnejad1.pdf | 1396/11/21 | 1392300 | دانلود |
