ساخت پروتئین نوترکیب FanC اشریشیا کلی انتروتوکسیژنیک مرتبط با اسهال گوساله

نوع مقاله: میکروبشناسی و ایمنی شناسی

نویسندگان

1 گروه میکروبیولوژی و ایمونولوژی، دانشکده دامپزشکی دانشگاه تهران، تهران، ایران

2 گروه ایمونولوژی، موسسه تحقیقات واکسن و سرم سازی رازی، کرج، ایران

3 پژوهشکده زیست فناوری و مهندسی زیستی، دانشگاه صنعتی اصفهان، اصفهان، ایران

چکیده

زمینه مطالعه: اسهال یکی از بیماری های شایع و مهمترین عامل مرگ و میر و تاخیر رشد در گوساله های زیر یک ماه است. در بین سویه های اشریشیا کلی انتروتوکسیژنیک که مسبب اسهال در گوساله های یک تا هفت روزه هستند، سویه‌های واجد فیمبریه F5 اهمیت زیادی دارند. بخش FanC فیمبریهF5  نقش اساسی در اتصال باکتری به یاخته های روده و آغاز بیماری داشته و پادتن های تولید شده بر علیه آن اثر حفاظتی دارند. هدف: ساخت و ارزیابی بیان پروتئین نوترکیب FanC اشریشیا کلی انتروتوکسیژنیک مرتبط با اسهال گوساله. روش کار: در این مطالعه پلاسمید پروکاریوتی ‌pET-28a حاوی قسمت fanC فیمبریه F5  طراحی و ساخته شد و تولید پروتئین حاصل از پلاسمید طراحی شده در باکتری (DE3) ا BL21 اثبات رسید. نتـایـج: حاصل این تحقیق ساخت پلاسمید پروکاریوتی pET-28a حامل ژن FanC و تهیه پروتئین خالص آن پس از بیان ژن در باکتری (DE3)اBL21 بود که با روش‌های تعیین توالی و وسترن بلاتینگ به اثبات رسید. نتیجـهگیـری نهایی: پروتئین FanC و سویه باکتری نوترکیب تولید شده را می‌توان در آزمون‌های تشخیصی و همچنین طراحی و تولید واکسن کلی باسیلوز گوساله به کار برد.

کلیدواژه‌ها


عنوان مقاله [English]

Production of Recombinant FanC of Enterotoxigenic Escherichia Coli Associated with Calf Diarrhea

نویسندگان [English]

  • Saeid Tabatabaei 1
  • Gholamraza Nikbakhat Brujeni 1
  • Majid Tebyanian 2
  • Khalil Zainel 1
  • Seyed Amir Hossein Jalali 3
1 1Department of Microbiology and Immunology, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran
2 2Department of Immunology, Razi Vaccine & Serum Research Institute, Karaj, Iran
3 3Department of Natural Resources, Institute of Biotechnology & Bioengineering, Isfahan University of Technology, Isfahan, Iran
چکیده [English]

BACKGROUND: Diarrhea is a common disease in the neonate calf which imposes significant economic burden on cattle industry around the world. During the first week after birth, Enterotoxigenic E.coli (ETEC) strains carrying F5 fimbria are one of the most important pathogens causing calf diarrhea. F5 fimbria is involved in early stage of pathogenesis and is responsible for attachment of bacteria to enterocytes; this attachment is mediated by FanC protein of F5 fimbria. Antibodies directed against F5 fimbriae play a significant role in prevention and control of the disease. Objectives: Evaluation and expression of recombinant expression of F5 Fimbriae of Enterotoxigenic Escherichia Coli associated with calf diarrhea. Methods: In the present study,  the fanC region of F5 fimbria was cloned in a pET28a plasmid. Results: The recombinant construct was confirmed by sequencing and protein production in Escherichia coli BL21 (DE3) was evaluated by western blotting procedure. Conclusions: Based on our findings, the recombinant FanC protein or the BL21 (DE3) strain are suitable candidates to develop an effective vaccine against calf colibacillosis or use in a diagnostic kit for F5+ ETEC.

کلیدواژه‌ها [English]

  • Cloning
  • Recombinant F5 protein
  • Bovine
  • Enterotoxigenic
  • E.coli
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