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| Construction and identification of single chain Fv fragment library used for ribosome display |
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Abstract The mice were immunized with Xanthomonas axonopodis pv. citri (Xac) and the mRNA of spleen cells from immunized mice were extracted. The variable fragment of heavy chain (VH) and light chain (VL) were amplified by RT-PCR and the linker (Gly4Ser)3 were used to ligate VH and VL. Then the single chain Fv fragment (scFv) library was constructed which would be used to screen positive scFvs by ribosome display. To analyze the diversity of the developed scFv library, the DNA of library was translated to Escherichia coli JM109, and the positive clones were selected by random and then sent to sequence. The results showed that the serum titer of immunized mice was about 2500×. The size of amplified VH were 350 bp and VL was 650 bp, and that of constructed scFv library DNA was 1200 bp. The scFv library were diversity and specific scFvs against Xac were selected from the constructed scFv library. It showed that the scFv library could be used to select positive scFvs against Xac by ribosome displaly.
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Received: 11 November 2008
Published: 27 April 2009
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