Construction of Candida albicans pRB895-SAP2-SC5314 With SAP2 High Expression and Its Effects on Adhesion

IF 2.6 4区医学 Q2 MEDICAL LABORATORY TECHNOLOGY

Journal of Clinical Laboratory Analysis Pub Date : 2024-12-27 DOI:10.1002/jcla.25144

Lan Xue, Lu Yang, Bingqian Zhao, Wenli Feng, Jing Yang, Yan Ma

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引用次数: 0

Abstract

Background

SAP2 is closely associated with the pathogenicity and drug resistance of Candida albicans (C. albicans). Our study aimed to construct C. albicans with SAP2 overexpression (pRB895-SAP2-SC5314) to explore the influence of SAP2 on the adhesion of C. albicans and predict the interaction between magnolol and Sap2 by molecular docking.

Methods

The recombinant plasmid pRB895-SAP2 with high SAP2 expression was prepared using a plasmid extraction kit and transformed into C. albicans strain SC5314 using an improved lithium acetate conversion method to construct PRB895-SAP2-SC5314. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) was performed to detect the expression of adhesion-related genes in the different strains. Molecular docking and visual analysis of magnolol and Sap2 were performed using the CB-DOCK2 platform.

Results

Compared with the control SC5341 and SC5341 transfected with pRB895, SAP2 expression was significantly higher in the pRB895-SAP2-SC5314 strain (p < 0.05). Based on the sequencing and mapping results, the pRB895-SAP2-SC5314 strain was successfully prepared. SAP2 overexpression significantly downregulated ALS1 expression (p < 0.05), whereas ALS3, TEC1, HOG1, PHR1, and TUP1 expression was downregulated in C. albicans (p < 0.05). The optimal docking result for magnolol and Sap2 protein was −8.1 kcal/mol of vina score, which was considered good docking.

Conclusions

SAP2 overexpression may strengthen the adhesion and pathogenicity of C. albicans, and magnolol may act as an Sap2 inhibitor that affects the adhesion of C. albicans.

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SAP2高表达白色念珠菌pRB895-SAP2-SC5314的构建及其对粘附的影响

背景：SAP2与白色念珠菌（C. albicans）的致病性和耐药性密切相关。本研究旨在构建SAP2过表达的白色念珠菌（pRB895-SAP2-SC5314），探讨SAP2对白色念珠菌粘附的影响，并通过分子对接预测厚朴酚与SAP2的相互作用。方法：采用质粒提取试剂盒制备SAP2高表达的重组质粒pRB895-SAP2，采用改进的醋酸锂转化法转化白色念珠菌SC5314，构建pRB895-SAP2 -SC5314。采用定量反转录聚合酶链反应（qRT-PCR）检测不同菌株中粘附相关基因的表达情况。利用CB-DOCK2平台对厚朴酚和Sap2进行分子对接和可视化分析。结果：与对照SC5341和转染pRB895的SC5341相比，pRB895-SAP2- sc5314菌株中SAP2的表达明显升高(p)。结论：SAP2过表达可增强白色念珠菌的粘附和致病性，厚朴酚可能作为SAP2抑制剂影响白色念珠菌的粘附。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Journal of Clinical Laboratory Analysis 医学-医学实验技术

CiteScore

5.60

自引率

7.40%

发文量

584

审稿时长

6-12 weeks

期刊介绍： Journal of Clinical Laboratory Analysis publishes original articles on newly developing modes of technology and laboratory assays, with emphasis on their application in current and future clinical laboratory testing. This includes reports from the following fields: immunochemistry and toxicology, hematology and hematopathology, immunopathology, molecular diagnostics, microbiology, genetic testing, immunohematology, and clinical chemistry.