华支睾吸虫副肌球蛋白功能区基因的序列分析及B细胞抗原表位预测  

Sequence analysis and B-cell antigenic epitope prediction of paramyosin functional region gene in Clonorchis sinensis

作　　者：邱阳元 马晓晓 常巧呈[1] 张艳[1] 王春仁[1] QIU Yangyuan;MA Xiaoxiao;CHANG Qiaocheng;ZHANG Yan;WANG Chunren(College of Animal Science and Veterinary Medicine,Heilongjiang Bayi Agricultural University,Daqing 163319,China)

机构地区：[1]黑龙江八一农垦大学动物科技学院

出　　处：《黑龙江畜牧兽医》2018年第21期117-119,262,263,共5页Heilongjiang Animal Science And veterinary Medicine

基　　金：国家重点研发计划项目(2017YFD0501300)

摘　　要：为了阐明华支睾吸虫副肌球蛋白（paramyosin,Pmy）的生物学特性,试验对其功能区的C段（PmyC）基因进行克隆、序列分析及B细胞抗原表位预测,采用TRIzol法提取华支睾吸虫成虫的总RNA,将其反转录成cDNA,再利用PCR方法对PmyC基因进行扩增,将其连接到pMD 18-T载体上进行克隆、测序,并利用DNAStar软件进行序列分析,同时利用生物学软件DNAStar 5. 0对B细胞抗原表位进行预测。结果表明：该虫体PmyC序列长度为903 bp,A＋T含量为52. 6%,与NCBI上的华支睾吸虫囊虫幼、麝后睾吸虫、卫氏并殖吸虫和日本血吸虫核苷酸序列的同源性分别为99. 4%、94. 0%、75. 0%和71. 1%。蛋白二级结构预测显示,PmyC蛋白主要由8个α-螺旋、2个β-折叠和3个β-转角构成。B细胞抗原表位预测显示,在该蛋白中有2个B细胞抗原表位。说明华支睾吸虫副肌球蛋白（CsPmy）具有成为抗原分子的潜能。In order to elucidate the biological characteristics of Clonorchis sinensis paramyosin（Pmy）,the C fragment functional（PmyC） gene of Clonorchis sinensis was cloned and sequenced,and the B-cell antigenic epitopes were predicted. The total RNA of adult Clonorchis sinensis was extracted by TRIzol method,and then c DNA was obtained by reverse transcription. PmyC gene was amplified by PCR,cloned into pMD18-T vector,sequenced and analyzed by DNASstar software. At the same time,the B-cell epitopes were predicted by biological software DNAStar5. 0. The results showed that the length of PmyC sequence of the parasite was 903 bp,the content of A＋T was 52. 6%. The homology of the nucleotide sequence of the parasite with the nucleotide sequences of Clororchis sinensis metacercaria Opisthorchis viverrini,Paragonimus westermani and Schistosoma japonicum on NCBI was 99. 4%,94. 0%,75. 0% and 71. 1%,respectively. Protein secondary structure prediction showed that PmyC protein was mainly composed of eight α-helix,two β-folds and three β-rotation. The prediction of B-cell antigen epitopes showed that there were two B-cell epitopes in the protein. The results indicated that the paramyosin of Clonorchis sinensis（CsPmy） had the potential to be an antigen molecule.

关 键 词：华支睾吸虫 副肌球蛋白 功能基因 序列分析 二级结构 表位预测 

分 类 号：S855.99[农业科学—临床兽医学] S852.735[农业科学—兽医学]