贵州农业科学2026，Vol.54Issue(8)：75-83,9.DOI:10.3969/j.issn.1001-3601.2026.08.009
豆伞滑刃线虫Bd-MIF-1基因的克隆与分析
Cloning and Analysis of Bd-MIF-1 Gene from Bursaphelenchus doui
摘要
Abstract
[Objective]The macrophage migration inhibitory factor(MIF)is an effector protein with an inhibitory role in host immune response.The MIF gene was cloned from Bursaphelenchus doui,and its expression characteristics were investigated to provide a reference for the function in the interaction process between B.doui and its hosts.[Method]The anterior-end specific cDNA library of B.doui was constructed by using the solid-phase hybridization method and using the anterior part,including esophageal gland,secretory pore and cerebral organ as the teaters and the posterior part,including intestine as the driver.The cDNA library was screened by reverse-northern blotting and sequencing technology.The full-length cDNA sequence of an MIF homologue was cloned by using RACE technology.The nucleotide sequence and amino acid sequence were analyzed by using the bioinformatics method.The gene copy number was identified by Southern blot method and the developmental expression pattern of MIF gene was detected by the real-time fluorescent quantitative PCR.[Result]The anterior-end specific cDNA library of B.doui was constructed successfully.The full-length cDNA sequence of Bd-MIF-1 gene(GenBank:PQ275932)was cloned from B.doui.The cDNA sequence of Bd-MIF-1 was 528 bp in length and contains 3 exons and 2 introns.The gene encoding had 114 amino acids.The relative molecular weight and theoretical isoelectric point were 12.16 kD and 6.22 respectively.The Bd-MIF-1 protein with a 4-Oxalocrotonate-Tautomerase superfamily structural domain had high homology with the amino acid sequence of Bursaphelenchus.The Southern blot analysis indicated that Bd-MIF-1 gene belonged to a single copy gene in B.doui genomes.The real-time fluorescent quantitative PCR result showed that the Bd-MIF-1 gene was expressed at all intart stages of Bursaphelenchus doui but there was no significant difference.[Conclusion]The Bd-MIF-1gene is successfully cloned from B.doui and its encoding protein contains a 4-Oxalocrotonate_Tautomerase superfamily domain.The encoding protein with a tauromerase activity may be involved in the parasitic process of B.doui.关键词
豆伞滑刃线虫/特异cDNA文库/巨噬细胞移动抑制因子/生物信息学/Southern杂交/表达分析Key words
Bursaphelenchus doui/specific cDNA library/macrophage migration inhibitory factor(MIF)/boin-formatics/Southern blot/expression analysis分类
农业科技引用本文复制引用
陈正伟,李莉,余婧,王仁刚,王自力,林世锋,朱秀,陈越男,吴沙沙..豆伞滑刃线虫Bd-MIF-1基因的克隆与分析[J].贵州农业科学,2026,54(8):75-83,9.基金项目
国家自然科学基金项目(32060342) （32060342）
贵州师范大学学术新苗基金项目(黔师新苗[2021]A13号) （黔师新苗[2021]A13号）
贵州省科技厅农业攻关项目(黔科合NY[2013]3041号) （黔科合NY[2013]3041号）
贵州省林业科研项目(黔林科合[2014]02号) （黔林科合[2014]02号）
中国烟草总公司贵州省公司科技项目(2023XM04) （2023XM04）
