Cloning and Expression of Cyclophilin from Platanus orientalis Pollens in Escherichia coli
Publish Year: 1391
نوع سند: مقاله ژورنالی
زبان: English
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شناسه ملی سند علمی:
JR_RBMB-1-1_005
تاریخ نمایه سازی: 10 شهریور 1400
Abstract:
Background: Allergy is a clinical disorder affecting the human population with wide geographical distribution. Platanus orientalis (P. orientalis) trees are planted in many countries and their pollen causes allergic reactions.
Cyclophilin has recently been identified as one of the most important allergens of P. orientalis pollen. We aimed to clone and purify this allergen in Escherichia coli for further studies and therapeutic and diagnostic purposes for allergy to P. orientalis.
Methods: RNA was extracted from P. orientalis. A full-length fragment encoding cyclophilin was prepared by polymerase chain reaction amplification of the first-strand cDNA synthesized from P. orientalis RNA. The cDNA was inserted into the pET۳۲b (+) vector, and the construct transformed into E. coli Top۱۰ and BL۲۱ cells. The expressed protein was purified by the CuSO۴ method.
Results: The cDNA for the cyclophilin of P. orientalis pollen was cloned, and a specific reactivity of recombinant cyclophin was confirmed by immunoblotting using sera from patients allergic to P. orientalis pollen.
Conclusion: The recombinant cyclophilin has a potential for immunologic assays for evaluation of allergy to P. orientalis pollen.
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Authors
Mojtaba Sankian
Immunology Research Center, Mashhad University of Medical Sciences, Mashhad, Iran
Fatemeh Vahedi
Razi Vaccine & Serum Research Institute, Mashhad, Iran
Nazanin Pazouki
Immunology Research Center, Mashhad University of Medical Sciences, Mashhad, Iran
Malihe Moghadam
Immunology Research Center, Mashhad University of Medical Sciences, Mashhad, Iran
Farahzad Jabbari Azad
Allergy Research Center, Mashhad University of Medical Sciences, Mashhad, Iran
Abdol-Reza Varasteh
Allergy Research Center, Mashhad University of Medical Sciences, Mashhad, Iran