摘要
【目的】分离和鉴定扩展莫尼茨绦虫(Moniezia expansa)电压门控钙通道β亚基(Cavβ)基因,预测蛋白二级结构和B细胞抗原表位,从而探讨Cavβ在寄生虫病治疗中所起的作用。【方法】构建扩展莫尼茨绦虫成虫cDNA文库,随机挑取重组阳性克隆进行测序,对部分序列进行引物步移法测序,获取全长cDNA序列;采用生物信息学方法对其对应的蛋白进行二级结构和B细胞抗原表位的预测。【结果】获得扩展莫尼茨绦虫Cavβ基因,全长946 bp,编码180个氨基酸,理论分子质量为19.788 8 kD,等电点为5.06,属于Cachannel B超家族。二级结构以无规则卷曲为主,结构预测存在4个可能的B细胞抗原表位。【结论】研究对于扩展莫尼茨绦虫Cavβ基因的获得和B细胞抗原表位的预测,为该基因功能的试验性鉴定工作奠定了基础。
【Objective】 The purpose of this project was to clone and identify the voltage-gated calcium channel beta subunit（Cavβ） gene and predict its secondary structure and B-cell epitopes,which might provide a theoretical basis for the pharmacotherapy of parasitic diseases.【Method】 A cDNA library was constructed from M.expansa adult stage.Clones were selected randomly from the cDNA library and sequenced with the method of primer-walking,and finally the full-length cDNA sequence was acquired.By using bioinformatices software and methods,the secondary structure and B-cell preponderant epitope were predicted.【Result】 The sequence of Cavβ gene was 946 base pairs and encoded polypeptide 180 amino acids with the predicted molecular mass of 19.788 8 kDa and theoretical pI of 5.06,which belong to the Ca_channel_B superfamily.Structure prediction indicated secondary structure were mainly random coil and there were 3 Protein kinase C phosphorylation site;1 Casein kinase II phosphorylation site;1 N-myristoylation site;1 Cell attachment sequence;4 possible B-cell epitopes,no transmembrane helix.【Conclusion】 The full-length cDNA encoding M.expansa Cavβ was obtained and B cell epitopes was predicted possible,which laid a basis for further functional study of this gene.
出处
《新疆农业科学》
CAS
CSCD
北大核心
2012年第4期735-742,共8页
Xinjiang Agricultural Sciences
基金
国家重点基础研究发展计划前期研究专项(2006CB708512)
新疆生产建设兵团杰出青年创新资金专项(2011CD005)
家畜疫病病原生物学国家重点实验室开放基金课题(SKLVEB2011KFKT008)
