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Construction of mouse scFv phage display library against tetrodotoxin and Establishment of TTX's Detection System

【摘要】:正In order to construct a single chain fragment variable(scFv) phage display library against tetrodotoxin(TTX),TTX-KLH complecation antigen was prepared,and used to immunize BALB/c mice.By using RT-PCR,the mouse heavy chain variable region genes(V_h) and light chain variable region genes(V_l) were amplified from speen of immunized mouse with TTX complecation antigen and subsequently assembled into scFv genes by overlap extension polymerase chain reaction(SOE-PCR).The resulting scFv genes were electrotransformed into E.coli TG1 and amplified with the co-infection of phage M13K07 to obtain phage display library.The capacity and titer of the resulting library were detected. After amplification with help phage,high-capacity phage antibody library was obtained.The mouse scFv library against TTX had been constructed successfully,suppling a tool which plays an important role in TTX poison therapy and studying of sodium channel.

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