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Updated: Sep 26, 2026
Functional Cloning Using a Xenopus Oocyte Expression System
Published on: January 30, 2016
Cloning, expression, crystallization, and preliminary X-ray analysis of recombinant mouse lipocalin-type
Daisuke Irikura1, Takashi Kumasaka, Masaki Yamamoto
1Core Research for Evolutional Science and Technology, Japan Science and Technology Corporation, 6-2-4 Furuedai, Suita, Osaka 565-0874.
Abstract:
Lipocalin-type prostaglandin D synthase is the key enzyme for the production of prostaglandin D(2), a potent endogenous somnogen, in the brain. We cloned, produced, and crystallized the native enzyme and selenomethionyl Cys(65)Ala mutants of the recombinant mouse protein by the hanging drop vapor-diffusion method with both malonate and citrate as precipitants. The native crystals obtained with malonate belong to orthorhombic space group P2(1)2(1)2(1) with lattice constants a = 46.2, b = 66.8, and c = 105.3 A. The selenomethionyl crystals obtained with citrate belong to orthorhombic space group C222(1) with lattice constants a = 45.5, b = 66.8, and c = 104.5 A. The native crystals diffracted beyond 2.1 A resolution.
