中国兽医科学2017，Vol.47Issue(5)：587-591,5.DOI:10.16656/j.issn.1673-4696.2017.05.009
肌旋毛虫谷氨酰胺酶基因的原核表达及其免疫荧光定位研究
Prokaryotic expression and immunofluorescent localization of glutaminase gene in Trichinella spiralis muscle larvae
摘要
Abstract
To analyze the enzymatic immunogenicity and reactivity of recombinant glutaminese decarboxylase TsGls protein of Trichinella spiralis muscle larvae,the complete sequence of TsGls gene was amplified by RT-PCR,cloned into prokaryotic expression plasmid pET-32a(+),transformed into Rosette(DE3),and then induced with IPTG.The polyclonal antibodies against the recombinant protein was prepared with the purified protein and then the immunofluorescent localization of TsGls in Trichinella spiralis muscle larvae was studied.The recombinant plasmid pET-32a(+)-TsGls was successfully constructed.The recombinant protein rTsGls was confirmed to be about 72 ku in size by SDS-PAGE.The high polyclonal antibody titer was up to 1.024 × 106,indicating good immunogenicity of rTsGls.Western-blot analysis showed high reactivity between rTsGls and larval extracts.Immunofluorescent showed that TsGls protein existed in the subcutaneous tissue of Trichinella spiralis.The above-mentioned results laid foundation for further study of glutamate-dependent acid mechanisms of Trichinellaspiralis.关键词
旋毛虫肌幼虫/谷氨酰胺酶/原核表达/免疫荧光定位Key words
Trichinella spiralis muscle larvae/glutaminase/prokaryotic expression/immunofluo-rescence localization分类
农业科技引用本文复制引用
赵祥,于洁,胡静,许静秀,韩彩霞,李晓云,宋铭忻..肌旋毛虫谷氨酰胺酶基因的原核表达及其免疫荧光定位研究[J].中国兽医科学,2017,47(5):587-591,5.基金项目
家畜疫病病原生物学国家重点实验室开放课题(SKLVEB2015KFKT010) （SKLVEB2015KFKT010）
国家自然科学基金项目(31172312) （31172312）
