construction of chimeric dual-chain avidin by tandem fusion of the related avidins串联融合建设嵌合双链式亲和素的抗生物素蛋白有关.pdf

Construction of Chimeric Dual-Chain Avidin by Tandem Fusion of the Related Avidins ¨ 1 1. 1.¤a ¨ ¨ 2¤b 3 Tiina A. Riihimaki , Sampo Kukkurainen , Suvi Varjonen , Jarno Horha , Thomas K. M. Nyholm , 1,2 ¨ 1,2 Markku S. Kulomaa , Vesa P. Hytonen * 1 Institute of Biomedical Technology, University of Tampere and Tampere University Hospital, Tampere, Finland, 2 Department of Biological and Environmental Science, ˚ ¨ ¨ ¨ ¨ University of Jyvaskyla, Jyvaskyla, Finland, 3 Department of Biochemistry and Pharmacy, Abo Akademi University, Turku, Finland Abstract Background: Avidin is a chicken egg-white protein with high affinity to vitamin H, also known as D-biotin. Many applications in life science research are based on this strong interaction. Avidin is a homotetrameric protein, which promotes its modification to symmetrical entities. Dual-chain avidin, a genetically engineered avidin form, has two circularly permuted chicken avidin monomers that are tandem-fused into one polypeptide chain. This form of avidin enables independent modification of the two domains, including the two biotin-binding pockets; however, decreased yields in protein production, compared to wt avidin, and complicated genetic manipulation of two highly similar DNA sequences in the tandem gene have limited the use of dual-chain avid