این سایت در حال حاضر پشتیبانی نمی شود و امکان دارد داده های نشریات بروز نباشند
Iranian Journal of Biotechnology، جلد ۱۳، شماره ۴، صفحات ۴۸-۵۲
عنوان فارسی
چکیده فارسی مقاله
کلیدواژه‌های فارسی مقاله
عنوان انگلیسی Heterologous Expression of Potato Virus Y Coat Protein, Isolate Pot187
چکیده انگلیسی مقاله Background: The advent of recombinant DNA technology has facilitated heterologous expression of proteins from various sources in different host systems including Escherichia coli. If a plant virus coat protein is expressed in the bacterium it can be used as the antigen for antibody preparation. Such a recombinant antigen preparation can be particularly useful where equipment such as ultracentrifuge is unavailable to purify virus particles to use as the antigen for conventional antibody preparation. Objective: Heterologous protein expression and purification of the full length Potato virus Y (PVY) coat protein (CP) from isolate pot187 (an affiliate of strain N) to be used as an antigen was the aim of the study. Materials and Methods: Reverse transcription Polymerase Chain Reaction (RT-PCR) was carried out to amplify an 801 bp fragment of the CP gene from PVY-infected potato leaves. The amplicon was cloned into pGEM-T Easy. The cloned fragment was restricted by BamHI + SacI and the purified fragment was cloned into the expression vector pET21a(+) which was restricted with the same enzymes. The generated plasmid was introduced into E. coli strain RosettaTM. The expression was induced with isopropyl-b-D-thiogalactopyranoside (IPTG) and its protein content was subjected to SDS-PAGE and western blotting. Results: SDS-PAGE analysis of protein from the induced bacteria showed a ~35 KDa protein corresponding to PVY CP. Expression of the recombinant protein was confirmed by anti-His anitibody. Conclusions: The full-length cDNA of PVY-CP was amplified from the infected potato leaves. The cDNA was heterologously expressed in E. coli. The produced recombinant CP can be used as an antigen to generate polyclonal antibody.
کلیدواژه‌های انگلیسی مقاله
نویسندگان مقاله نعمت سخندان بشیر | sokhandan bashir
department of plant protection, university of tabriz, tabriz, iran
سازمان اصلی تایید شده: دانشگاه تبریز (Tabriz university)

مهین poorsmaile |
department of plant protection, university of tabriz, tabriz, iran
سازمان اصلی تایید شده: دانشگاه تبریز (Tabriz university)

محمد حاجی زاده |
department of plant protection, university of kurdistan, sanandaj, iran

نشانی اینترنتی http://www.ijbiotech.com/article_10910_340696a94f9325fc97e535906745a6dd.pdf
فایل مقاله فایلی برای مقاله ذخیره نشده است
کد مقاله (doi)
زبان مقاله منتشر شده en
موضوعات مقاله منتشر شده
نوع مقاله منتشر شده
برگشت به: صفحه اول پایگاه   |   نسخه مرتبط   |   نشریه مرتبط   |   فهرست نشریات