中国农业科学2013，Vol.46Issue(10)：2175-2182,8.DOI:10.3864/j.issn.0578-1752.2013.10.024
莫氏巴贝斯虫滑动体组件蛋白基因gap45、gap50、myo A和m/c的克隆表达与多克隆抗体制备
Cloning and Expressing Glideosome Associated Protein Genes of Babesia motasi and Preparation of Their Polyclonal Antibodies
摘要
Abstract
[Objective] To prepare polyclonal antibodies of component genes from glideosome,gap45,gap50,myo A,and mlc were cloned and expressed in prokaryotic system.[Method] Rapid amplification of cDNA end (RACE) was used to amplify the full lengths of gap45,gap50,myo A,and mlc.The partial or complete open reading frames (ORF) of those genes were further inserted into prokaryotic expression vector and then transformed into BL21 (DE3) pLys to express by inducing with IPTG.The purified fusion proteins,with MagneGSTTM Protein Purification System,were injected into rabbits to produce polyclonal antibodies.Specificities and titers of the polyclonal antibodies were determined by ELISA assay.[Result] The full-length cDNAs of gap45,gap50,myo A,and mlc,were 664,1391,2538,773 bp and open reading frames (ORF) were 567,1194,2514,and 606 bp in size.The polyclonal antibodies prepared had good specificities and the titers were more than 1∶25600.[Conclusion] In the present study,the full length of four component genes of B.motasi glideosome were amplified and their polyclonal antibodies were obtained,which will provide important biological materials for screening and identifying candidate antigens of vaccination and diagnosis,drug targets as well studying invasion mechanism of Babesia in the future.关键词
巴贝斯虫/RACE/滑动体/克隆/表达/多克隆抗体Key words
Babesia/ RACE/ glideosome/ clone/ expression/ polyclonal antibody引用本文复制引用
王锦明,刘爱红,任巧云,马米玲,刘志杰,李安岩,李有全,赵帅阳,张浩浩..莫氏巴贝斯虫滑动体组件蛋白基因gap45、gap50、myo A和m/c的克隆表达与多克隆抗体制备[J].中国农业科学,2013,46(10):2175-2182,8.基金项目
国家自然科学基金项目(31072130,30800820,30972182,31001061)、甘肃省重点项目(0801NKDA033,1002NKDA035)、"973"项目(2010CB530206)、"948"项目(2010-S04)、国家肉牛牦牛产业体系项目(CARS-38)、科技部合作专项、欧盟EPIZONE(FOOD-CT-2006-016236)、ASFRISK(№211691)、ARBOZOONET(№211757)、PIROVAC(KBBE-3-245145) （31072130,30800820,30972182,31001061）
