یافته‌‌های اخیر در زمینه توکسین‌های NetB و TpeL جهت تولید واکسن علیه بیماری انتریت نکروتیک طیور

نوع مقاله : مقاله مروری

نویسندگان

بخش تحقیق و تولید واکسن های بی هوازی، آزمایشگاه تخصصی تحقیقات کلستریدیا، موسسه واکسن و سرم سازی رازی، سازمان ترویج آموزش توسعه کشاورزی، کرج، ایران

چکیده

کلستریدیوم شامل طیف گسترده‌ای از باکتری‌های میله‌ای شکل بی هوازی گرم مثبت و اسپورزا است که دارای 203 گونه بوده و بر اساس چهار توکسین اصلی یوتا (iA)، آلفا (cpa)، بتا (cpb) و اپسیلون، به پنج تایپ (A، B، C، D، E) طبقه‌بندی می‌شود. این باکتری‌ها یکی از مهم‌ترین عوامل بیماری‌زایی در حیوانات هستند. توکسین‌های متنوع این باکتری نقش مهمی در بیماری‌زایی دارند که از جمله جدیدترین آن‌ها می‌توان به NetB و TpeL اشاره نمود. این توکسین‌ها در ایجاد بیماری‌های متنوعی از جمله انتریت نکروتیک نقش دارند. در این مقاله مروری بر مطالعات انجام یافته بر روی کلیات توکسین‌های NetB و TpeL ارائه می‌گردد. همچنین پیشرفت‌های اخیر جهت تولید واکسن‌های نسل جدید را ارائه و تجزیه و تحلیل‌هایی که توسط دانشمندان با استفاده از نرم‌افزارهای بیوانفورماتیک روی توالی‌های tpeL و netB برای تولید پروتئین نوترکیب انجام یافته، مورد بررسی قرار می‌گیرد. در این راستا محققین در آزمایشات مختلف ژن فیوژن را توسط PCR سنتز نموده و پلاسمید نوترکیب را در باکتری E. coli ترانسفورم کرده‌اند. پروتئین فیوژن حاصله توسط نیکل(Ni-NTA) خالص شده و جهت تأیید از تکنیک‌های SDS-PAGE و وسترن‌بلات استفاده گردیده و جهت سنجش ایمنی، آزمایش خنثی‌سازی سرم در خرگوش استفاده شده است. همچنین در مطالعات دیگر موتانت‌هایی با فعالیت سایتوتوکسیک کمتر توسط کیت به صورت site-directed mutagenesis طراحی شده و با توالی‌سازی تأیید شدند. نتایج مطالعات آنان حاکی از آن است که واکسن‌های نسل جدید می‌توانند به عنوان کاندید واکسن علیه انتریت نکروتیک طیور مورد استفاده قرار گیرند. 

کلیدواژه‌ها

عنوان مقاله [English]

Recently acquired for NetB and TpeL toxins for vaccine production to protect against avian necrotic enteritis

نویسندگان [English]

  • L. Abdolmohammadi Khiav
  • R. Pilechian Langroodi
  • A. Paradise
Department of Anaerobic Vaccine Research and Production, Specialized Clostridia Research Laboratory, Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization, Karaj, Iran
چکیده [English]

Clostridium is a broad genus of rod-shaped, anaerobic, gram-positive and spore-forming bacteria. It has 203 species and is classified into five isotypes (A, B, C, D, and E) based on four major toxins, iota (iA), alpha (cpa), beta (cpb) and epsilon (etx). These bacteria are one of the most important causes of disease in animals. Various toxins of these bacteria play an important role in pathogenesis. Newly discovered toxin such as NetB and TpeL could be mentioned. These toxins are involved in a variety of diseases, including necrotic enteritis. This article presents an overview of studies on NetB and TpeL toxins. Furthermore recent advances had been provided in the production of new generation vaccines. In this article, we demonstrated that the sequences of tpeL and netB were investigated using bioinformatics software for recombinant protein production. Different regions of the gene were selected and evaluated for fusion protein synthesis. The fusion gene was synthesized by PCR and the recombinant plasmid was transformed into E. coli. Then the fusion protein was purified by NI-NTA and used for confirmation by SDS-PAGE and Western blotting. Serum neutralization test was done to evaluate of antibody in rabbits. Mutants with less cytotoxic activity were also designed by site-directed mutagenesis kits and confirmed by sequencing. The results of this study suggested that new-generation vaccines could be used as a candidate vaccine against avian necrotic enteritis.

کلیدواژه‌ها [English]

  • TpeL
  • NetB
  • Clostridium perfringens
  • new-generation vaccines
  • bioinformatics

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تحقیقات دامپزشکی و فرآورده‌های بیولوژیک
دوره 34، شماره 1
فروردین 1400
صفحه 26-37

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سابقه مقاله

  • تاریخ دریافت: 18 آذر 1398
  • تاریخ پذیرش: 30 بهمن 1398

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