曼氏迭宫绦虫annexinB8的生物信息学分析和基因克隆

发布时间：2019-05-17 14:30

【摘要】：目的通过生物信息学预测曼氏迭宫绦虫annexinB8(SmannexinB8)的生物学特征,及潜在功能和结构,并且进行基因克隆,为下一步SmannexinB8参与宿主免疫调节研究提供依据。方法通过NCBI的ORF finder工具对SmannexinB8的开放阅读框进行分析.利用ExPASy网站进行蛋白的物理化学参数、信号肽、跨膜螺旋、潜在分子生物学功能的预测,通过NCBI/BLAST。对蛋白保守功能域进行检测。不同物种的annexin序列从NCBI网站获取,并利用Vector NTI suit 8.0和TreeView软件进行分析。利用SWISS-MODEL网站和SPDBV 4.10软件分析SmannexinB8蛋白的三维空间结构。此外。对SmannexinB8基因进行扩增,并克隆到原核表达载体pET-28a(+)。结果 SmannexinB8是一个全长基因,编码347个氨基酸。蛋白由4个典型的annexin重复结构域组成,序列当中没有信号肽,是一个稳定的可溶性蛋白分子。三维空间立体结构分析结果显示SmannexinB8是一个保守的蛋白。SmannexinB8与多房棘球绦虫、口膜壳绦虫、细粒棘球绦虫、华支睾吸虫以及人类的annexin基因的同源性分别是68%,67%。65%,46%和40%。分子进化分析显示SmannexinB8与绦虫属的亲源性最近,而与其他物种,如吸虫、哺乳动物亲源性较远。结论 SmannexinB8可能具有抑制磷脂酶A2的活性,促进细胞融合、参与调节免疫反应和离子通道形成的功能,可能在参与宿主免疫调节中起到关键性的作用。
[Abstract]:Objective to predict the biological characteristics, potential function and structure of annexinB8 (SmannexinB8) by bioinformatics, and to clone the gene, so as to provide the basis for the next step of SmannexinB8 to participate in the study of host immunomodulation. Methods the open reading box of SmannexinB8 was analyzed by ORF finder tool of NCBI. The physicochemical parameters, signal peptides, transmembrane helix and potential molecular biological functions of proteins were predicted by ExPASy website. NCBI/BLAST. was used to predict the physicochemical parameters, signal peptides, transmembrane helix and potential molecular biological functions of proteins. The conserved domain of protein was detected. The annexin sequences of different species were obtained from NCBI website and analyzed by Vector NTI suit 8.0and TreeView software. SWISS-MODEL website and SPDBV 4.10 software were used to analyze the three-dimensional spatial structure of SmannexinB8 protein. In addition. The SmannexinB8 gene was amplified and cloned into prokaryotic expression vector pET-28a (). Results SmannexinB8 was a full-length gene encoding 347 amino acids. The protein is composed of four typical annexin repeat domains, and there is no signal peptide in the sequence, so it is a stable soluble protein molecule. The results of three-dimensional spatial structure analysis showed that SmannexinB8 was a conserved protein .Smannexin B8. The homology of annexin gene with Echinococcus multilocularis, Echinococcus stomata, Echinococcus granulosus, Clonorchis sinensis and human was 68% and 67%, respectively. 46% and 40%. Molecular evolution analysis showed that SmannexinB8 had the closest affinity with tapeworm, but far from other species, such as trematodes and mammals. Conclusion SmannexinB8 may inhibit the activity of phospholipase A2, promote cell fusion, participate in the regulation of immune response and ion channel formation, and may play a key role in host immune regulation.
【作者单位】：海南医学院病原生物学教研室;海南省人民医院检验科;
【基金】：国家自然科学基金课题(81260254) 海南省自然科学基金课题(814289)
【分类号】：R383.3

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