Cloning and expression in Escherichia coli of cDNA encoding house dust mite allergen Der f 3, serine protease from Dermatophagoides farinae

FEBS Letters

Yasuhara, Takaomi¹  Yuuki, Toshifumi¹  Okumura, Yasushi¹  Nishiyama, Chiharu¹ 

[1] Central Research Laboratories, Asahi Breweries Ltd., 2-13-1 Ohmori-kita, Ohta-ku, Tokyo 143, Japan

关键词: Der f 3;    Allergen;    Serine protease;    cDNA;    Sequence;   

DOI  :  10.1016/0014-5793(95)01291-5

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Der f 3 is one of the allergens produced by house dust mite Dermatophagoides farinae showing serine protease activity. Based on its amino acid sequence, a cDNA clone encoding Der f 3 was isolated from a cDNA library of D. farinae. Sequencing analysis of the clone revealed the presence of an open reading frame of 780 bp, which encodes a mature protein of 232 amino acids with 27 amino acids of pre-pro sequence at the N-terminus. When proDer f 3 was produced in Escherichia coli as a fused protein with glutathione-S-transferase, the fused protein was accumulated as inclusion bodies. The protein purified with 8 M urea and glutathione-affinity column chromatography, however, did not show protease activity. When an arginine residue was introduced at the C-terminus of the pro-region in place of threonine, removal of the pro-region to produce an active mature protease was observed. The specificity and the activity of this recombinant protease were almost the same as those of native Der f 3.

【 授权许可】

Unknown