中国农业大学学报2024，Vol.29Issue(10)：199-207,9.DOI:10.11841/j.issn.1007-4333.2024.10.17
捻转血矛线虫blister-5基因序列特征及表达特性分析
Analysis of sequence characteristics and expression properties of blister-5 in Haemonchus contortus
摘要
Abstract
The study was to identify the expression properties of the Kunitz-type protein BLI-5 in Haemonchus contortus.Firstly,analysis of its gene structure,conserved domain,evolutionary relationships and protein three-dimensional structure using bioinformatics software were conducted;Secondly,the relative transcript abundance of Hc-bli-5 and Hc-bli-4 was analyzed based on WormBase database at each developmental stage of H.contortus;Subsequently,a prokaryotic expression vector to obtain recombinant protein and prepared polyclonal antibodies were constructed,while tissue distribution of the protein candidates in the worm was analyzed by the immunolocalization assay.The results revealed that:1)The Hc-bli-5 gene has a total of 2 088 bp,and consisted of 4 exons and 3 introns.Hc-BLI-5 has a total of 195 amino acids,with 132-192 aa forming a Kunitz-type super family domain,which is a serine proteinase inhibitor.2)Phylogenetic tree showed that BLI-5 has a close evolutionary relationship in both parasitic and free-living nematodes,and the evolutionary relationship between H.contortus and Haemonchus placei is most closely aligned,clustering together on the same minor branch of the phylogenetic tree;3)Comparative analysis of protein structures indicated that Hc-BLI-5 and Ce-BLI-5 shared certain similarities,especially in the Kunitz domain.4)Transcript abundance analysis at each developmental stage showed that the expression trend of Hc-bli-5 and Hc-bli-4 overlapped,suggesting that they may interact with each other and participate in common biological process.5)Immunolocalization showed that BLI-5 was distinctly expressed at the cuticle and muscle tissue of worm.In summary,Hc-bli-5 may interact with Hc-bli-4 and participate in the molting process of H.contortus.This study clarified the sequence characteristics and expression properties of Hc-bli-5,providing a theoretical basis for exploring its involvement in the molting process and mechanism in H.contortus.关键词
捻转血矛线虫/蜕皮/Hc-bli-5/序列特征/表达特性Key words
Haemonchus contortus/molting/Hc-bli-5/sequence characteristics/expression property分类
畜牧业引用本文复制引用
汤洪宁,吴飞,项继忠,童丹妮,杜振东,陈学秋,马光旭,杨怡..捻转血矛线虫blister-5基因序列特征及表达特性分析[J].中国农业大学学报,2024,29(10):199-207,9.基金项目
国家自然科学基金项目(32202829,32172877) （32202829,32172877）
浙江省自然科学基金项目(LZ22C180003,LQ23C180006) （LZ22C180003,LQ23C180006）
