PMID: 18724711Aug 30, 2008Paper

Cloning of foot-and-mouth disease virus integrin receptor beta1 subunit and antibody production to its ligand-binding domain

Sheng wu gong cheng xue bao = Chinese journal of biotechnology
Ping DuXiangtao Liu

Abstract

We produced beta1 gene which is about 2400 bp by reverse transcription polymerase chain reaction (RT-PCR) from bovine trachea, reclaimed and purified, then cloned the amplified fragment to pGEM-T easy vector, confirmed by sequencing. The immune-dominant epitope of beta1 gene was chosen by computer analysis and then syncretized ligand-binding domain from 346 bp to 843 bp of ecytoplasm with six histidine, expressed LBD protein massly in E. coli BL21 (DE3), and identified by SDS-PAGE. The fusion protein was purified with Ni-NTA affinity chromatography and immunized New Zealand rabbits preparing of its polyclonal antibody, the specific antibody titer was above 1:12,800 detected by indirect ELISA, the result of Western blot showed that this antibody could be recognized by LBD fusion protein.

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