Construction of the recombinant Listeria monocytogenes mutant expressing green fluorescent protein

A recombinant Listeria monocyogenes mutant expressing green fluorescent protein (GFP) gene was constructed by homologous recombination. The target gene gfp was fused to the actA gene fragment downstream of its promoter and signal sequence. PCR amplification and restriction digestion of the PCR produ...

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Main Authors: KE Chun-lin, SONG Hou-hui, JIANG Ling-li, ZHANG Gui-zhi, FANG Wei-huan
Format: Article
Language:English
Published: Zhejiang University Press 2005-09-01
Series:浙江大学学报. 农业与生命科学版
Subjects:
Online Access:https://www.academax.com/doi/10.3785/1008-9209.2005.05.0638
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author KE Chun-lin
SONG Hou-hui
JIANG Ling-li
ZHANG Gui-zhi
FANG Wei-huan
author_facet KE Chun-lin
SONG Hou-hui
JIANG Ling-li
ZHANG Gui-zhi
FANG Wei-huan
author_sort KE Chun-lin
collection DOAJ
description A recombinant Listeria monocyogenes mutant expressing green fluorescent protein (GFP) gene was constructed by homologous recombination. The target gene gfp was fused to the actA gene fragment downstream of its promoter and signal sequence. PCR amplification and restriction digestion of the PCR product confirmed the insertional fusion. The reporter gene was expressed as shown by fluorescent microscopy and SDS-PAGE/western blotting. The recombinant strain exhibited reduced invasiveness to the in vitro cultured cells as well as decreased virulence to the chicken embryos and mice. The strain has potential applications in microbe-host interaction studies in the medical or veterinary medical fields and in food microbiological studies.
format Article
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institution DOAJ
issn 1008-9209
2097-5155
language English
publishDate 2005-09-01
publisher Zhejiang University Press
record_format Article
series 浙江大学学报. 农业与生命科学版
spelling doaj-art-c3f087f3f0944d3e96ffcffe96d301e22025-08-20T03:16:04ZengZhejiang University Press浙江大学学报. 农业与生命科学版1008-92092097-51552005-09-013163864410.3785/1008-9209.2005.05.063810089209Construction of the recombinant Listeria monocytogenes mutant expressing green fluorescent proteinKE Chun-linSONG Hou-huiJIANG Ling-liZHANG Gui-zhiFANG Wei-huanA recombinant Listeria monocyogenes mutant expressing green fluorescent protein (GFP) gene was constructed by homologous recombination. The target gene gfp was fused to the actA gene fragment downstream of its promoter and signal sequence. PCR amplification and restriction digestion of the PCR product confirmed the insertional fusion. The reporter gene was expressed as shown by fluorescent microscopy and SDS-PAGE/western blotting. The recombinant strain exhibited reduced invasiveness to the in vitro cultured cells as well as decreased virulence to the chicken embryos and mice. The strain has potential applications in microbe-host interaction studies in the medical or veterinary medical fields and in food microbiological studies.https://www.academax.com/doi/10.3785/1008-9209.2005.05.0638<italic>Listeria monocytogenes</italic>homologous recombinationgreen fluorescent proteinheterologous gene expression
spellingShingle KE Chun-lin
SONG Hou-hui
JIANG Ling-li
ZHANG Gui-zhi
FANG Wei-huan
Construction of the recombinant Listeria monocytogenes mutant expressing green fluorescent protein
浙江大学学报. 农业与生命科学版
<italic>Listeria monocytogenes</italic>
homologous recombination
green fluorescent protein
heterologous gene expression
title Construction of the recombinant Listeria monocytogenes mutant expressing green fluorescent protein
title_full Construction of the recombinant Listeria monocytogenes mutant expressing green fluorescent protein
title_fullStr Construction of the recombinant Listeria monocytogenes mutant expressing green fluorescent protein
title_full_unstemmed Construction of the recombinant Listeria monocytogenes mutant expressing green fluorescent protein
title_short Construction of the recombinant Listeria monocytogenes mutant expressing green fluorescent protein
title_sort construction of the recombinant listeria monocytogenes mutant expressing green fluorescent protein
topic <italic>Listeria monocytogenes</italic>
homologous recombination
green fluorescent protein
heterologous gene expression
url https://www.academax.com/doi/10.3785/1008-9209.2005.05.0638
work_keys_str_mv AT kechunlin constructionoftherecombinantlisteriamonocytogenesmutantexpressinggreenfluorescentprotein
AT songhouhui constructionoftherecombinantlisteriamonocytogenesmutantexpressinggreenfluorescentprotein
AT jianglingli constructionoftherecombinantlisteriamonocytogenesmutantexpressinggreenfluorescentprotein
AT zhangguizhi constructionoftherecombinantlisteriamonocytogenesmutantexpressinggreenfluorescentprotein
AT fangweihuan constructionoftherecombinantlisteriamonocytogenesmutantexpressinggreenfluorescentprotein