关键词: 鸭肠炎病毒')">鸭肠炎病毒, UL53基因, B细胞表位, 原核表达

Abstract:

【Objective】 Applying the strategy to select the main antigenic domains of DEV-UL53 gene and carry out multiparameter B cell epitope prediction of DEV-truncated UL53 gene, to express efficiently the DEV-truncated UL53 gene and analyze the antigenicity of this fussion protein by Western blot. 【Method】 The bioinformatics software DNAStar Protean was used to predict the main antigenic domains of gK protein coded by DEV-UL53 gene, then the truncated UL53 gene corresponding the main antigenic domains was selected and used to predict the secondary structure, flexibility domains, surface possibility with DNAStar software and predict hydrophilicity as well as transmembrane domains on-line, meanwhile clone, subclone as well as prokaryotic expression of the truncated UL53 gene, also the antigenicity of the fussion protein was detected by Western blot. 【Result】 The B cell epitopes of gK coded by DEV-truncated UL53 gene most likely distribute in Ala20—Leu25, Ser40—Met47, Leu68—Ile78, Val124—Phe128, Ile129—Tyr134, Asp176—Ile178. The recombinant expression plasmid was transformed into BL21 and expressed under the induction of IPTG. Western blot analysis indicated that polyclonal rabbit antiserum against DEV had specific reaction with the fussion protein. 【Conclusion】 DEV-truncated UL53 gene was efficiently expressed in prokaryotic system and also the gK fussion protein has good reactionogenicity, which provided useful data for research of the UL53 gene, gK protein and development of new generational vaccine and diagnostic reagent.

Key words: duck enteritis virus')">duck enteritis virus, UL53 gene, B cell epitope, prokaryotic expression