Abstract:In order to clone and express the gene of hookworm platelet inhibitor from Ancylostoma ceylanicum (AceHPI) and analyze its biological characteristics, the specific primers were designed referring to the published sequence of AcaHPI from Ancylostoma caninum. The gene was amplified by RT-RCR, and the PCR product was cloned into pET-28a vector for expression by induction of IPTG. Meanwhile, the homology and bioinformatics analysis of amino acid sequences encoded by the gene was conducted by online software. The results showed that the open reading frame of AceHPI gene was 603 bp in length, encoding a protein of 200 amino acids, and its sequence homology with AcaHPI was 91%. The recombinant expression plasmid pET28a-AceHPI was constructed and expressed, and a soluble fusion protein with a molecular mass of about 26 kDa was obtained. Predictive analysis indicated that the protein was hydrophobin and in the first 17 amino acids was signal peptide sequences, without transmembrane domain. In this study, AceHPI gene was successfully cloned and expressed, and its encoded amino acids were analyzed by bioinformatics, laying a foundation for the further study on the mechanism of AceHPI’s role in the infected host.
黄岳, 刘云秋, 颜欣欣, 杭建雄, 冉荣坤, 孙永祥, 李国清. 锡兰钩虫HPI基因的原核表达与生物信息学分析[J]. 中国人兽共患病学报, 2019, 35(8): 706-710.
HUANG Yue, LIU Yun-qiu, YAN Xin-xin, HANG Jian-xiong, RAN Rong-kun, SUN Yong-xiang, LI Guo-qing. Prokaryotic expression and bioinformatics analysis of HPI gene of Ancylostoma ceylanicum. Chinese Journal of Zoonoses, 2019, 35(8): 706-710.
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