Construction of Ribosome Display Single Chain Antibody Library of Foot and Mouth Disease

China Biotechnology

2011, Vol. 31

Issue (7): 98-103 DOI:

Construction of Ribosome Display Single Chain Antibody Library of Foot and Mouth Disease

YANG Shuang, MA Jun-wu, ZHOU Guang-qing, LIN Mi, FENG Xia, LIU Tao, DAI Peng

Lanzhou Veterinary Research Institute, Chinese Academy of Agriculture Science, State Key Laboratory of Veterinary Etiological Biology, National Foot-and-Mouth Disease Reference Laboratory, Lanzhou 730046, China

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Abstract

Objective: To construct a rabbit source high-capacity and good-diversity ribosome display single chain antibody (scFv) library of FMDV. Methods: The isolated spleen cells from rabbit immuned by the FMDV Virus,and extracted RNA for amplifying rabbit VH gene, VL gene and Ck gene as the spacer by RT-PCR,and then connect VH-VL gene by overlapping extension PCR (referred to as SOE-PCR). After that,the elements for ribosome display such as T7 promoter and ribosome binding site were introduced after SOE products were amplified. Finally, the ribosome display scFv library template was constructed in vitro. Moreover, the ribosome display templates were connected to pMD18-T vector into E.coli DH5α, and the positive clones were picked sequencing to identify the ScFv assembly. Results: We successfully constructed a rabbit source ribosome display scFv library whose capacity is 8.21 × 10¹³ of FMDV. Conclusion: The rabbit-derived high-capacity ribosome display antibody library can be a good platform for screening specific foot and mouth disease, and lay a good experimental basis for the measuring test of FMDV.

Key words： Ribosome display technology Overlapping extension PCR scFv library FMDV

Received: 26 January 2011 Published: 25 July 2011

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YANG Shuang, MA Jun-wu, ZHOU Guang-qing, LIN Mi, FENG Xia, LIU Tao, DAI Peng. Construction of Ribosome Display Single Chain Antibody Library of Foot and Mouth Disease. China Biotechnology, 2011, 31(7): 98-103.

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https://manu60.magtech.com.cn/biotech/ OR https://manu60.magtech.com.cn/biotech/Y2011/V31/I7/98

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