کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
8421404 | 1545924 | 2015 | 22 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Construction of bacterial ghosts for transfer and expression of a chimeric hepatitis C virus gene in macrophages
ترجمه فارسی عنوان
ساخت ارواح باکتریایی برای انتقال و بیان یک ژن ویروسی هپاتیت سی در ماکروفاژها
دانلود مقاله + سفارش ترجمه
دانلود مقاله ISI انگلیسی
رایگان برای ایرانیان
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
بیوتکنولوژی یا زیستفناوری
چکیده انگلیسی
The bacterial ghost (BG) production is a field of biotechnology for applications in vaccine and drug delivery. We assessed the capacity of BG for delivery of a recombinant gene encoded for both cell mediated and antibody dependent epitopes of hepatitis C virus (HCV) into murine macrophages. Escherichia coli (E. coli) cells were transformed with the lysis plasmid (pHH43). To produce chimeric gene, NS3 (non-structural protein 3) and core regions of HCV genome were fused together by splicing by overlap extension (SOEing) PCR and were cloned into plasmid pEGFP-C1. Bacterial ghosts were loaded with recombinant pEGFP-C1 and then were transferred to murine macrophages (RAW 264.7). To investigate plasmid transfection and chimeric mRNA transcription, fluorescent microscopy and RT-PCR were used. In vitro studies indicated that bacterial ghosts loaded with pEGFP-C1 plasmid were efficiently taken up by murine macrophages and indicated a high transfection rate (62%), as shown by fluorescent microscopy. RT-PCR from extracted intracellular mRNAs for chimeric Core-NS3 gene showed a specific 607Â bp fragment of the gene. The sequence analysis of purified PCR products demonstrated the expected unique mRNA sequence. We constructed a chimeric HCV gene containing both cell mediated and antibody dependent epitopes with a significant expression in murine macrophages delivered by bacterial ghost.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Microbiological Methods - Volume 119, December 2015, Pages 228-232
Journal: Journal of Microbiological Methods - Volume 119, December 2015, Pages 228-232
نویسندگان
M.R. Miri, A. Behzad-Behbahani, M. Fardaei, A. Farhadi, Y. Talebkhan, M. Mohammadi, M. Tayebinia, F. Farokhinejad, P. Alavi, M. Fanian, F. Zare, J. Saberzade, N. Nikouyan, M.A. Okhovat, R. Ranjbaran, G. Rafiei Dehbidi, S. Naderi,