Knocking out FLO8 gene of Candida glabrata and its effect on EPA family

Objective To construct a FLO8 gene knockout strain of Candida glabrata and analyze the effect of FLO8 knockout on the expression of EPA family in Candida glabrata. Methods By fusion PCR technology, the knockout components were constructed with the genomic DNA of Candida glabrata ATCC2001 strain and...

Full description

Saved in:
Bibliographic Details
Main Author: ZHAO Juntao, YUAN Jie, LIU Jinyan, CHEN Kezhi, XIANG Mingjie
Format: Article
Language:zho
Published: Editorial Office of Journal of Diagnostics Concepts & Practice 2024-08-01
Series:Zhenduanxue lilun yu shijian
Subjects:
Online Access:https://www.qk.sjtu.edu.cn/jdcp/fileup/1671-2870/PDF/1732171224734-490173967.pdf
Tags: Add Tag
No Tags, Be the first to tag this record!
_version_ 1846161386180182016
author ZHAO Juntao, YUAN Jie, LIU Jinyan, CHEN Kezhi, XIANG Mingjie
author_facet ZHAO Juntao, YUAN Jie, LIU Jinyan, CHEN Kezhi, XIANG Mingjie
author_sort ZHAO Juntao, YUAN Jie, LIU Jinyan, CHEN Kezhi, XIANG Mingjie
collection DOAJ
description Objective To construct a FLO8 gene knockout strain of Candida glabrata and analyze the effect of FLO8 knockout on the expression of EPA family in Candida glabrata. Methods By fusion PCR technology, the knockout components were constructed with the genomic DNA of Candida glabrata ATCC2001 strain and the plasmid DNA with screening marker NAT as templates. The knockout components were transfected into Candida glabrataATCC2001 by lithium acetate transfection method to obtainflo8△strain. The expression of EPA1, EPA6 and EPA7 genes were detected by real-time quantitative PCR. Results The FLO8 gene knockout strain of Candida glabrata was efficiently constructed. The expression levels of EPA1, EPA6 and EPA7genes in flo8△strain were significantly lower than those in ATCC2001 strain (all P<0.001). Conclusions The gene knockout method permits rapid, easy and highly efficient generation of homozygous knockout mutations in Candida glabrata. The knockout of FLO8 gene reduced the expression of EPA family in Candida glabrata, which laid a foundation for further study on its virulence mechanism.
format Article
id doaj-art-e5daf76390c8402190184ea1c5c245d8
institution Kabale University
issn 1671-2870
language zho
publishDate 2024-08-01
publisher Editorial Office of Journal of Diagnostics Concepts & Practice
record_format Article
series Zhenduanxue lilun yu shijian
spelling doaj-art-e5daf76390c8402190184ea1c5c245d82024-11-21T09:06:48ZzhoEditorial Office of Journal of Diagnostics Concepts & PracticeZhenduanxue lilun yu shijian1671-28702024-08-01230439840410.16150/j.1671-2870.2024.04.008Knocking out FLO8 gene of Candida glabrata and its effect on EPA familyZHAO Juntao, YUAN Jie, LIU Jinyan, CHEN Kezhi, XIANG Mingjie01. Department of Clinical Laboratory, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China;2. Radioimmunology and ClinicalLaboratory, Ruijin HospitalLuwan Branch, Shanghai Jiao Tong University School of Medicine, Shanghai 200020, ChinaObjective To construct a FLO8 gene knockout strain of Candida glabrata and analyze the effect of FLO8 knockout on the expression of EPA family in Candida glabrata. Methods By fusion PCR technology, the knockout components were constructed with the genomic DNA of Candida glabrata ATCC2001 strain and the plasmid DNA with screening marker NAT as templates. The knockout components were transfected into Candida glabrataATCC2001 by lithium acetate transfection method to obtainflo8△strain. The expression of EPA1, EPA6 and EPA7 genes were detected by real-time quantitative PCR. Results The FLO8 gene knockout strain of Candida glabrata was efficiently constructed. The expression levels of EPA1, EPA6 and EPA7genes in flo8△strain were significantly lower than those in ATCC2001 strain (all P<0.001). Conclusions The gene knockout method permits rapid, easy and highly efficient generation of homozygous knockout mutations in Candida glabrata. The knockout of FLO8 gene reduced the expression of EPA family in Candida glabrata, which laid a foundation for further study on its virulence mechanism.https://www.qk.sjtu.edu.cn/jdcp/fileup/1671-2870/PDF/1732171224734-490173967.pdf|candida glabrata|gene knockout|adhesin|flo8 gene|epithelial adhesin family
spellingShingle ZHAO Juntao, YUAN Jie, LIU Jinyan, CHEN Kezhi, XIANG Mingjie
Knocking out FLO8 gene of Candida glabrata and its effect on EPA family
Zhenduanxue lilun yu shijian
|candida glabrata|gene knockout|adhesin|flo8 gene|epithelial adhesin family
title Knocking out FLO8 gene of Candida glabrata and its effect on EPA family
title_full Knocking out FLO8 gene of Candida glabrata and its effect on EPA family
title_fullStr Knocking out FLO8 gene of Candida glabrata and its effect on EPA family
title_full_unstemmed Knocking out FLO8 gene of Candida glabrata and its effect on EPA family
title_short Knocking out FLO8 gene of Candida glabrata and its effect on EPA family
title_sort knocking out flo8 gene of candida glabrata and its effect on epa family
topic |candida glabrata|gene knockout|adhesin|flo8 gene|epithelial adhesin family
url https://www.qk.sjtu.edu.cn/jdcp/fileup/1671-2870/PDF/1732171224734-490173967.pdf
work_keys_str_mv AT zhaojuntaoyuanjieliujinyanchenkezhixiangmingjie knockingoutflo8geneofcandidaglabrataanditseffectonepafamily