چاپ صفحه |  صفحه نخست سامانه |  نویسندگان |  اطلاعات تفضیلی |  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 | 87356 |
| عنوان فارسی مقاله | بیان دمین اتصال گیرنده SARS-CoV-2 (RBD) در سیستم پروکاریوتی |
| عنوان لاتین مقاله | Expression of SARS-CoV-2 receptor binding domain (RBD) in prokaryotic system |
| نوع ارائه | پوستر |
| عنوان کنگره / همایش | biomedical congress |
| نوع کنگره / همایش | بین المللی |
| کشور محل برگزاری کنگره/ همایش | |
| شهر محل برگزاری کنگره/ همایش | کنگره مجازی و آنلاین است |
| سال انتشار/ ارائه شمسی | 1401 |
| سال انتشار/ارائه میلادی | 2022 |
| تاریخ شمسی شروع و خاتمه کنگره/همایش | 1401/08/19 الی 1401/08/28 |
| آدرس لینک مقاله/ همایش در شبکه اینترنت | https://www.icbcongress.com/2022/Archives?o=303&lang=fa |
| آدرس علمی (Affiliation) نویسنده متقاضی | Drug Applied Research Center, Tabriz University of Medical Sciences, Tabriz, Iran |
| نویسنده | نفر چندم مقاله |
|---|---|
| سمانه جهاندار لاشکی | اول |
| صفر فرج نیا | دوم |
| مرتضی میلانی | چهارم |
| عنوان | متن |
|---|---|
| کلمات کلیدی | Receptor binding domain (RBD), protein expression, E. coli BL21, COVID-19, SARS-CoV-2. |
| خلاصه مقاله | Introduction: The receptor binding domain (RBD) is a part of the SARS-CoV-2 S protein, which binds to the host receptor and facilitates viral entry. RBD expressed in eukaryotic cells cannot meet the high expression yield and cost requirements for therapeutics and vaccines. There have been numerous studies suggesting that the RBD expressed by E. coli may induce protective immunity. Furthermore, RBD expressed in E. coli has been used for worldwide research purposes as a cost-effective antigen. In this study, we expressed RBD in the E. coli BL21 strain and evaluated the purified protein by SDS page and ELISA. Methods: The plasmid and the sequence coding for RBD were treated with restriction enzymes. The RBD fragment was ligated into pET28a (+) using T4 DNA ligase. Then the expression construct was transformed into E. coli BL21, cultured in LB media, and induced with isopropyl b-D-1-thiogalactoside. A 15% SDS-PAGE was used to analyze the expression. For purification, the cells were lysed by sonication and recombinant RBD was purified by the NiNTA column. Finally, the recombinant RBD was refolded by the gradual elimination of urea by dialysis. The quality of purified RBD was assessed by SDS-PAGE analysis and ELISA. Results: To produce a large amount of antigen for the biopanning process, the DNA sequence of the RBD protein of SARS-CoV-2 spike protein was synthesized and cloned into the PET-28a vector. Results of PCR analysis and DNA sequencing confirmed the recombinant PET-28a sequence at the appropriate size of ~ 4000 bp as shown in supplementary data 1. The recombinant protein was expressed in E. coli strain BL21, purified by NINTA column, and refolded by dialysis. The SDS-PAGE results showed 95% purification of the RBD protein. To ensure the accurate antigenicity and function of the RBD protein an ELISA test on pooled COVID-19 serum samples was performed and the results revealed a high binding affinity of the serum antibodies to the RBD protein. Conclusion: In conclusion, we expressed high yields of SARS-CoV-2 receptor binding domain protein in E. coli strain BL21. Our results indicate that expressed RBD is functional and can induce immune responses |
| نام فایل | تاریخ درج فایل | اندازه فایل | دانلود |
|---|---|---|---|
| abstract.pdf | 1404/01/30 | 204514 | دانلود |
| ICB2022 .pdf | 1404/01/30 | 1513411 | دانلود |
