Optimization of Expression and Extraction of Toxocara canis Recombinant C-Type Lectin Protein in Escherichia coli BL21 (DE3) Article Swipe
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· 2021
· Open Access
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· DOI: https://doi.org/10.22059/jvr.2020.287488.2966
زمینۀ مطالعه: توکسوکاریازیس یک بیماری مهم زئونوز است که توسط نوزاد مرحله دوم توکسوکارا کنیس و توکسوکارا کتی ایجاد میشود. در خیلی از کشورهای معتدل توکسوکاریازیس رایجترین عفونت کرمی است و سبب بروز عوارض شدید میشود. لکتین نوع سی از محصولات تولیدی در مرحله نوزادی این انگل است. این پروتئین در واکنشهای ایمنی ازجمله ارسال سیگنال سلولهای ایمنی در مهرهداران، فعال کردن ایمنی ذاتی در مهرهداران و بیمهرگان و القا هموستاز دخالت دارد.هدف: مطالعه حاضر با هدف بهینهسازی تولید پروتئین نوترکیب لکتین نوع سی انگل توکسوکارا کنیس و بررسی خواص آنتیژنی (پادگنی) آن صورت گرفته است.روشکار: توالی نوکلئوتیدی مرجع مربوط به لکتین نوع سی(CTL) انگل توکسوکارا کنیس از بانک ژن استخراج شد. سپس پلاسمید pET32a حاوی توالی مورد نظر توسط شرکت GENERAY سنتز گردید. پلاسمید نوترکیب به باکتری اشریشیا کلای سویه BL21(DE3) منتقل شد. بیان این پروتئین با استفاده از روشهای SDS-PAGE و داتبلات بررسی و با سرم انسانی مثبت، مورد تأیید قرار گرفت.نتایج: مطالعه حاضر نشان داد که بهینهسازی شرایط برای دستیابی به بیشترین میزان تولید پروتئین با انتخاب باکتری اشریشیا کلای سویه BL21(DE3)، دمای 37 درجه سانتیگراد پس از القا و غلظت 1 میلیمولارIPTG حاصل گردید. پس از بهینهسازی پروتئین نوترکیب با غلظت 6/0±1160میکروگرم بر میلیلیتر استخراج شد. وزن مولکولی پروتئین حاصل 42 کیلودالتون بود. پاساژ پلاسمید نوترکیب قبل از القا، در باکتری اشریشیا کلای سویه DH5α باعث افزایش معنیدار بیان پروتئین گردید. نتایج ارزیابی بهینهسازی شرایط، با روشهای SDS-PAGE و داتبلات نشان داد که بیشترین میزان تولید پروتئین نوترکیب لکتین نوع سی انگل توکسوکارا کنیس در شرایط بهینهسازی حاصل شده است.نتیجهگیری نهایی: جهت تولید هر پروتئین نوترکیب بهویژه در مقادیر زیاد و بهمنظور مصارف تجاری لازم است که بهینهسازی مخصوص آن پروتئین، انجام شود و دقیقا براساس شرایط بهینهسازی تعیینشده، تولید پروتئین نوترکیب قابل انجام است. نتایج مطالعه حاضر نشان داد که شرایط بهینه جهت تولید پروتئین نوترکیب لکتین نوع سی، انتخاب باکتری اشریشیا کلای سویه BL21(DE3)، دمای 37 درجه سانتیگراد و مدت زمان 4 ساعت پس از القا است که با حفظ ویژگیهای ایمنیزایی نیز همراه است. بهطورکلی میتوان چنین نتیجهگیری کرد که سویه باکتری انتخاب شده و همچنین دما و زمان القا بر میزان بیان و تولید پروتئینهای نوترکیب مانند لکتین نوع سی انگل توکسوکارا کنیس تأثیر مستقیم دارد.
Related Topics
- Type
- article
- Language
- en
- Landing Page
- https://jvr.ut.ac.ir/article_83328_52c9a1d7038b3e0da8c7e1a21c32c238.pdf
- OA Status
- green
- Cited By
- 1
- Related Works
- 20
- OpenAlex ID
- https://openalex.org/W3200819086
Raw OpenAlex JSON
- OpenAlex ID
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https://openalex.org/W3200819086Canonical identifier for this work in OpenAlex
- DOI
-
https://doi.org/10.22059/jvr.2020.287488.2966Digital Object Identifier
- Title
-
Optimization of Expression and Extraction of Toxocara canis Recombinant C-Type Lectin Protein in Escherichia coli BL21 (DE3)Work title
- Type
-
articleOpenAlex work type
- Language
-
enPrimary language
- Publication year
-
2021Year of publication
- Publication date
-
2021-06-22Full publication date if available
- Authors
-
Mahsa Shahbakhsh, Fateme Jalousian, Seyed Hossein Hosseini, Parviz Shayan, Abdorreza Naser MoghadasiList of authors in order
- Landing page
-
https://jvr.ut.ac.ir/article_83328_52c9a1d7038b3e0da8c7e1a21c32c238.pdfPublisher landing page
- Open access
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YesWhether a free full text is available
- OA status
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greenOpen access status per OpenAlex
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https://jvr.ut.ac.ir/article_83328_52c9a1d7038b3e0da8c7e1a21c32c238.pdfDirect OA link when available
- Concepts
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Recombinant DNA, Lectin, Escherichia coli, Toxocara canis, Biology, Microbiology, C-type lectin, Affinity chromatography, Chemistry, Molecular biology, Biochemistry, Immunology, Helminths, Enzyme, GeneTop concepts (fields/topics) attached by OpenAlex
- Cited by
-
1Total citation count in OpenAlex
- Citations by year (recent)
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2022: 1Per-year citation counts (last 5 years)
- Related works (count)
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20Other works algorithmically related by OpenAlex
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| abstract_inverted_index.سی(CTL) | 103 |
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| abstract_inverted_index.ارسال | 54 |
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| abstract_inverted_index.همراه | 329 |
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| abstract_inverted_index.گرفته | 94 |
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| abstract_inverted_index.است.نتیجهگیری | 253 |
| cited_by_percentile_year.max | 94 |
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