空肠弯曲菌cdtA、cdtB、cdtC基因的克隆、序列分析及B细胞表位预测  

Clone,sequence analysis and prediction of secondary structure and B cell epitope of cdtA,cdtB,cdtC gene of campylobacter jejuni

作　　者：刘卫卫[1] 白欣立[2] 李震中[1] 张军峰[3] 李鑫[1] 赵子春[1] 李春岩[1] 

机构地区：[1]河北医科大学第二医院神经内科,石家庄050000 [2]河北医科大学第二医院儿科,石家庄050000 [3]中国人民解放军二六零医院神经内科

出　　处：《脑与神经疾病杂志》2010年第5期324-332,共9页Journal of Brain and Nervous Diseases

基　　金：国家自然科学基金(81072481);国家高技术研究发展计划(863计划2006AA02A208)

摘　　要：目的预测和分析空肠弯曲菌(CJ)细胞扩张毒素(CDT)cdtA、cdtB、cdtC蛋白的二级结构和B细胞抗原表位,为CJ疫苗研制提供基础资料。方法 PCR扩增cdtA、cdtB、cdtC基因,克隆入pGEM-T载体并测序,对重组质粒测序后进行生物信息学分析。以cdtA、cdtB、cdtC基因推导的氨基酸序列为基础,采用Chou-Fasman法、Ganmier-Robson法和Karplus-Schulz法预测该蛋白二级结构;按Kyte-Doolittle方案、Emini方案和JamesonWolf方案预测其B细胞抗原表位。结果克隆的cdtA、cdtB、cdtC基因全长分别为874bp、913bp、711bp。发现cdtA的840-32bp序列完全与标准株NCTC11168的1-807bp序列一致,cdtB的841-42bp序列完全与标准株NCTC11168的1-798bp序列一致,cdtC的667-96bp序列完全与标准株NCTC11168的1-570bp序列一致。cdtA第11～44和54～90区段,cdtB第5～70、78～112、137～187区段,cdtC第119～140和145～193区段及其附近可能是CJ-cdtA、cdtB、cdtC蛋白的B细胞表位优势区。结论成功扩增空肠弯曲菌O∶19菌株cdtA、cdtB、cdtC基因,测序表明与标准株序列高度同源,已有文献证明cdtA、cdtB、cdtC基因在不同CJ菌株中高度保守,符合疫苗候选抗原的基本特征;预测出cdtA、cdtB、cdtC蛋白的B细胞抗原表位优势区,这将为CJ新型疫苗的设计和单克隆抗体的筛选等方面的研究提供重要信息。Objective Predicting the secondary structure and B cell antigenic epitopes of cytolethal distending toxin(CDT)protein of campylobacter jejuni to provide grounding data means of preparing CJ vaccine.Methods The cdtA,cdtB,cdtC fragment was amplified from CJ O:19 chromo somal DNA by PCR.It was cloned into pGEM-T,sequenced and compared with international standard strains NCTC11168 in the GenBank.The cdtA,cdtB,cdtC gene of CJ were collected to predict the secondary structure by using methods of Chou-Fasman,Garnier-Robson and Karplus-Schulz,the hydrophilicity,surface probability and antigenic index of deduced cdtA,cdtB,cdtC by Kyte-Doolittle,Emini and Jameson-Wolf were analyzed respectively.The B cell antigenic epitopes for deduced cdtA,cdtB,cdtC were predicted by synthesizing these methods.Results The cdtA,cdtB,cdtC fragments were composed of 874,913,711 base pairs.And their nucleotide homology with Cj NCTC11168 strains on the GenBank were 100%.The predominant epitopes were probably in the region of 11～44 and 54～90 of cdtA,5～70,78～112 and 137～187 of cdtB,119～140 and 145～193 of cdtC.Conclusion The cdtA,cdtB,cdtC fragments are successfully amplified and their homology is high compared with otherspecies.We predict the secondary structure and B cell antigenic epitopes of cdtA,cdtB,cdtC.It will lay the basis for designing new generation vaccine of CJ and screeing the monoclonal antibody.

关 键 词：骨髓间充质干细胞 神经元 诱导分化 神经节苷脂园 空肠弯曲菌 克隆 序列分析 B细胞抗原表位 

分 类 号：R378[医药卫生—病原生物学]