Recombinant Allergen Production in E. coli

Methods Mol Biol. 2017:1592:23-45. doi: 10.1007/978-1-4939-6925-8_3.

Abstract

Recombinant protein allergens have been used in allergy studies, allergy diagnosis, and epitope mapping. Messenger RNAs (mRNAs) are isolated from tissues of interest for complementary DNA (cDNA) library construction. Subsequently, the allergen gene is amplified by polymerase chain reaction (PCR) and sequenced. The amplified gene is then cloned into an expression vector, expressed in Escherichia coli cells, and purified from the cell lysate. This chapter describes the protocols for recombinant allergen production.

Keywords: Allergy; Food allergen; Recombinant allergen.

MeSH terms

  • Allergens / genetics*
  • Cloning, Molecular / methods
  • DNA, Complementary / genetics
  • Escherichia coli / genetics*
  • RNA, Messenger / genetics
  • Recombinant Proteins / genetics*

Substances

  • Allergens
  • DNA, Complementary
  • RNA, Messenger
  • Recombinant Proteins