Recombinant Allergen Production in E. coli.
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.
School of Exercise and Nutritional Sciences, San Diego State University, 308 ENS Bldg., 5500 Campanile Drive, San Diego, CA, 92182-7251, USA.
Changqi
C
Liu
School of Science, Mathematics and Computing, Albany State University, Albany, GA, 31707, USA.
LeAnna N
LN
Willison
Department of Nutrition, Food and Exercise Sciences, Florida State University, 420 Sandels Bldg., 120 Convocation Way, Tallahassee, FL, 32306-1493, USA. ssathe@fsu.edu.
Shridhar K
SK
Sathe
D000485:Allergens / Q000235:genetics*; D003001:Cloning, Molecular / Q000379:methods; D018076:DNA, Complementary / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics*
D000485:Allergens; D018076:DNA, Complementary; D012333:RNA, Messenger; D011994:Recombinant Proteins
Allergy; Food allergen; Recombinant allergen