巨型住肉孢子虫烯醇酶基因克隆与表达

doi:10.11843/j.issn.0366-6964.2019.03.023

Abstract

Abstract:

Sarcocystis gigantea is a globally ubiquitous pathogen that infects sheep. The formation of cyst in striated muscle induces a largely waste of commercial mutton which causes livestock husbandry losses. In this study, we tried to explore the special antigens for serological diagnosis of sarcocystosis. We screened a series of candidate antigens in S. gigantea through Western blot and mass spectrum identification. Whereafter, Genome Walking technology was applied to acquire two flanking sequences of these antigens. Ultimately, purify recombinant proteins was used to evaluate diagnostic value by bioinformatics analysis and Western blot. Results were as follows:Enolase of S. gigantea was finally supposed to a candidate diagnostic antigen, named SgENO. Subsequently, 1 181 bp expressive sequence of SgENO was cloned, and recombinant protein rSgENO was obtained. Nevertheless, it was found to have high homology with enolase amino acids of other Apicomplexa (71%-92.1%), rSgENO could cross react with Toxoplasma gondii and Neospora caninum positive serum. We cloned and expressed the S. gigantea enolase, and the recombinant protein had a cross reaction with other parasitic protozoa positive serum, which made it an irrelevant antigen for Sarcocystis specific diagnose. However, this protein may be a candidate antigen for vaccines.

CLC Number:

S852.723

WANG Xianmei, WU Jiajia, LI Yi, LIU Qun, LIU Jing. Clone and Expression of Enolase in Sarcocystis gigantea[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(3): 670-676.

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Clone and Expression of Enolase in Sarcocystis gigantea

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