畜牧兽医学报2017，Vol.48Issue(11)：2173-2180,8.DOI:10.11843/j.issn.0366-6964.2017.11.019
柔嫩艾美耳球虫多肽:N-乙酰氨基半乳糖转移酶2的基因表达及转录动态分析
Expression Profiling of Polypeptide:N-acetylgalactosaminyltransferase T2 during Eimeria tenella Developmental Cycle
摘要
Abstract
O-linked glycosylation is a form of protein posttranslational modifications that occurs in the Golgi apparatus of eukaryotes,and archaea as well as bacteria.Polypeptide:N-acetylgalactosaminyltransferase 2 (ppGalNAc-T2,EC 2.4.1.41) is the first key enzyme in the pathway of O-glycan biosynthesis.Whole genome sequencing of Eimeria spp has ever revealed that these protozoa possess the pathways for O-glycan synthesis and O-linked glycosylation of proteins.For exploring the profiles of glycosylation in the global proteome of Eimeria tenella during developmental cycle,and the potential druggable target of EtppGalNAc-T2,we have cloned and recombinantly expressed the EtppGalNAc-T2 gene according to its predicted coding sequence deposited in EuPathDB,and evaluated its transcriptional profiling in different stages of life cycle using qRT-PCR method.The results showed that the ORF of EtppGalNAc-T2 is 1 983 bp in full length,and can be expressed in E.coli Transetta (DE3) with recombinant vector EtppGalNAc-T2-pGEX-6p-1.qRT-PCR results showed that the transcriptions of EtppGalNAc-T2 are significantly varied among different developmental stages,that is,the highest in E.tenella sporozoite,and nearly no-detectable transcription in sporulated occyst.Briefly,these results confirmcd O-glycan synthesis process,and may play roles in the regulation of developmental conversion in E.tenella.关键词
柔嫩艾美耳球虫/ppGalNAc-T2/基因克隆/表达差异/qRT-PCRKey words
Eimeria tenella/ppGalNAc-T2/gene cloning/expression profile/qRT-PCR分类
农业科技引用本文复制引用
刘晓丽,龚振兴,马雪婷,曲自刚,韩政岚,刘保红,袁如意,杨孝朴,蔡建平..柔嫩艾美耳球虫多肽:N-乙酰氨基半乳糖转移酶2的基因表达及转录动态分析[J].畜牧兽医学报,2017,48(11):2173-2180,8.基金项目
国家自然科学基金(31272554) （31272554）
中国农业科学院农业科技创新工程专项(2014-LVRI-09) （2014-LVRI-09）
