title: Cloning and expression of cyclophilin from Platanus orientalis pollens in Escherichia coli.
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 pET32b (+) vector, and the construct transformed into E. coli Top10 and BL21 cells. The expressed protein was purified by the CuSO4 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.
  
  
  CONCLUSIONS
  The recombinant cyclophilin has a potential for immunologic assays for evaluation of allergy to P. orientalis pollen.
authors.affiliation: Immunology Research Center, Mashhad University of Medical Sciences, Mashhad, Iran.
authors.forename: Mojtaba
authors.initials: M
authors.lastname: Sankian
authors.affiliation: Razi Vaccine & Serum Research Institute, Mashhad, Iran.
authors.forename: Fatemeh
authors.initials: F
authors.lastname: Vahedi
authors.affiliation: Immunology Research Center, Mashhad University of Medical Sciences, Mashhad, Iran.
authors.forename: Nazanin
authors.initials: N
authors.lastname: Pazouki
authors.affiliation: Immunology Research Center, Mashhad University of Medical Sciences, Mashhad, Iran.
authors.forename: Malihe
authors.initials: M
authors.lastname: Moghadam
authors.affiliation: Allergy Research Center, Mashhad University of Medical Sciences, Mashhad, Iran.
authors.forename: Farahzad
authors.initials: F
authors.lastname: Jabbari Azad
authors.affiliation: Allergy Research Center, Mashhad University of Medical Sciences, Mashhad, Iran.
authors.forename: Abdol-Reza
authors.initials: AR
authors.lastname: Varasteh
keywords: Allergy; Recombinant allergen; Cyclophilin; Cloning; Escherichia coli; Platanus orientalis; Pollen