摘要
为扩增环形泰勒虫微线体一棒状体蛋白基因的侧翼序列，从感染环形泰勒虫的淋巴细胞系中提取RNA，反转录合成第1链cDNA。以该cDNA为模板，利用高效热不对称交互PCR法（hiTAIL-PCR）进行扩增，将目的片段连接到pGEM—TEasy载体并进行测序。结果显示，获得了环形泰勒虫微线体一棒状体蛋白基因的5’端侧翼序列，该序列编码894个氨基酸。生物信息学分析结果表明，该蛋白具有信号肽，信号肽剪切部位位于第19～20个氨基酸残基之间，很可能是一种分泌蛋白；经TMHMM2．0预测，该蛋白在第875～892位氨基酸残基部位具有典型的跨膜结构；Motifscan预测的结果显示，环形泰勒虫微线体一棒状体蛋白存在2处N-糖基化位点（153NLSF、333NESM）和15处CK2蛋白激酶磷酸化位点；在该蛋白第500～650位氨基酸残基部分具有较高的抗原性，推测可能是该蛋白与其他蛋白发生相互作用的区域。
In order to amplify the flanking sequence of microneme-rhoptry protein gene from Theileria annulata, total RNA was extracted from the lymphocytes which was infected by T. annulata and the first strand cDNA was synthesized by reverse transcription. Using cDNA as template, flanking fragments were amplified by high-efficiency thermal asymmetric interlaced PCR（hiTAIL-PCR）. Sequence of the flanking fragment was cloned into pGEM-T Easy vector,and sequenced. Sequencing results showed that the 5~ flan- king sequence of the gene was obtained. Bioinformatics analysis revealed that the protein encoded by the se- quence had a signal peptide,which suggests it to be a secreted protein, and a transmembrane structure at 875--892 amino acid residues. In addition there were 2 N-glycosylation sites（153NLSF,333NESM） and 15 pro- tein kinase CK2 phosphorylation sites. And the 500- 650 amino acid residues had the highest antigenicity,which was supposed to be a protein-protein interaction region.
出处
《中国兽医科学》
CAS
CSCD
北大核心
2014年第1期1-6,共6页
Chinese Veterinary Science
基金
国家自然科学基金资助项目(31272556,31101621)
农业部肉牛牦牛项目(CARS-38)
国家重点基础研究发展计划(973)项目(2010CB530206)
国家引进国际先进农业科学技术计划(948)项目(2010-S04)
家畜疫病病原生物学国家重点实验室项目
欧盟EPIZONE项目(FOOD-CT-2006-016236)
ASFRISK项目(211691)
AR-BOZOONET项目(211757)
PIROVAC项目(KBBE-3-245145)
