新疆农业大学学报2026，Vol.49Issue(2)：99-108,10.DOI:10.20088/j.cnki.jxau.2026.02.001
球孢白僵菌XJBB-01株Pr1丝氨酸蛋白酶高效表达菌株的构建及其对拉合尔钝缘蜱毒力测定
Construction of High-Efficiency Expression Strain of Pr1 Serine Protease from Beauveria bassiana XJBB-01 and Its Determination of Virulence against Ornithodoros lahorensis
摘要
Abstract
This study utilized Agrobacterium tumefaciens-mediated transformation and hygromycin re-sistance screening to introduce the Pr1 serine protease CDEP2 gene into Beauveria bassiana strain XJBB-01.Enzyme activity assays were conducted to obtain a high-efficiency expression strain of Pr1 serine protease in Beauveria bassiana.The virulence of strain Pr1 serine protease activity before and after transformation,as well as the spore suspension concentration of 1×107 CFU/mL,were measured at different stages of Or-nithodoros lahorensis.The results demonstrated that the engineered Beauveria bassiana strains Pr1-1 and Pr1-2 were successfully obtained through hygromycin resistance screening and PCR identification.Pheno-typic analysis of the two engineered strains revealed significantly higher colony growth rates and spore pro-duction compared to the XJBB-01 strain(P<0.05).Among them,the colony growth rates of Pr1-1 and Pr1-2 were 9.27 mm/d and 8.87 mm/d,respectively,which were 0.77 mm/d and 1.17 mm/d higher than those of strain XJBB-01.The spore production rates were 4.58×106 spores/cm2 and 5.22×106 spores/cm2,respectively,representing increases of 0.51×106 spores/cm2 and 1.15×106 spores/cm2 compared to strain XJBB-01.The enzyme activities of XJBB-01 strain and Pr1-1,Pr1-2 strains were meas-ured,with Pr1-1 and Pr1-2 showing enzyme activities of 34.13 U/mL and 35.58 U/mL,respectively,repre-senting significant increases of 5.46 U/mL and 6.91 U/mL compared to XJBB-01 strain(P<0.05).At 25℃and a spore suspension concentration of 1×107 CFU/mL,strains Pr1-1 and Pr1-2 exhibited signifi-cantly lower hatching rates of Ornithodoros lahorensis eggs compared to strain XJBB-01,with reductions of 23.3%and 26.6%,respectively.The mortality rates of nymphs increased by 17.4%and 20.7%,while the mortality rates of adult ticks rose by 16.7%and 23.3%.The median lethal time of larval ticks was accel-erated by 2.3 d and 2.5 d,respectively,while that of adult ticks was accelerated by 2.6 d and 3.3 d,show-ing significant differences(P<0.05).关键词
球孢白僵菌/Pr1丝氨酸蛋白酶/遗传转化/拉合尔钝缘蜱/毒力测定Key words
Beauveria bassiana/Pr1 serine protease/genetic transformation/Ornithodoros lahorensis/virulence assay分类
农业科技引用本文复制引用
马思楠,周雨芊,张伟,刘丹丹,韩佳依,王永青,李龙涛,黄景仁杰,张泽田,李欣如,黄洁..球孢白僵菌XJBB-01株Pr1丝氨酸蛋白酶高效表达菌株的构建及其对拉合尔钝缘蜱毒力测定[J].新疆农业大学学报,2026,49(2):99-108,10.基金项目
国家自然科学基金项目(32060803) （32060803）
新疆维吾尔自治区天山英才创新团队项目(2023TSYCTD0008) （2023TSYCTD0008）
新疆维吾尔自治区兽医学特色学科和新疆草食动物新药研究与创制重点实验室开放课题(XJCDVM-HDRC-S202412) （XJCDVM-HDRC-S202412）
新疆农业大学大学生创新项目(dxscx2025113) （dxscx2025113）
