Construction of a bacteriophage-derived recombinase system in Bacillus licheniformis for gene deletion

Abstract Bacillus licheniformis and its related strains have found extensive applications in diverse industries, agriculture, and medicine. However, the current breeding methods for this strain primarily rely on natural screening and traditional mutagenesis. The limited availability of efficient gen...
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Gespeichert in:

Autor*in:	Fang Xue [verfasserIn] Xufan Ma [verfasserIn] Cheng Luo [verfasserIn] Dongliang Li [verfasserIn] Guiyang Shi [verfasserIn] Youran Li [verfasserIn]

Format:	E-Artikel
Sprache:	Englisch

Erschienen:	2023

Schlagwörter:	Bacillus licheniformis Homologous recombination Recombinase RecT

Übergeordnetes Werk:	In: AMB Express - SpringerOpen, 2011, 13(2023), 1, Seite 15
Übergeordnetes Werk:	volume:13 ; year:2023 ; number:1 ; pages:15

Links:	Link aufrufen Link aufrufen Link aufrufen Journal toc

DOI / URN:	10.1186/s13568-023-01589-w

Katalog-ID:	DOAJ092860338

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520			\|a Abstract Bacillus licheniformis and its related strains have found extensive applications in diverse industries, agriculture, and medicine. However, the current breeding methods for this strain primarily rely on natural screening and traditional mutagenesis. The limited availability of efficient genetic engineering tools, particularly recombination techniques, has hindered further advancements in its applications. In this study, we conducted a comprehensive investigation to identify and characterize a recombinase, RecT, derived from a Bacillus phage. Remarkably, the recombinase exhibited a 105-fold enhancement in the recombination efficiency of the strain. To facilitate genome editing, we developed a system based on the conditional expression of RecT using a rhamnose-inducible promoter (Prha). The efficacy of this system was evaluated by deleting the amyL gene, which encodes an α-amylase. Our findings revealed that the induction time and concentration of rhamnose, along with the generation time of the strain, significantly influenced the editing efficiency. Optimal conditions for genome editing were determined as follows: the wild-type strain was initially transformed with the genome editing plasmid, followed by cultivation and induction with 1.5% rhamnose for 8 h. Subsequently, the strain was further cultured for an additional 24 h, equivalent to approximately three generations. Consequently, the recombination efficiency reached an impressive 16.67%. This study represents a significant advancement in enhancing the recombination efficiency of B. licheniformis through the utilization of a RecT-based recombination system. Moreover, it provides a highly effective genome editing tool for genetic engineering applications in this strain.
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allfieldsSound	10.1186/s13568-023-01589-w doi (DE-627)DOAJ092860338 (DE-599)DOAJ760eeb63251e4ccca3e36bf7af14f284 DE-627 ger DE-627 rakwb eng TP248.13-248.65 QR1-502 Fang Xue verfasserin aut Construction of a bacteriophage-derived recombinase system in Bacillus licheniformis for gene deletion 2023 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Abstract Bacillus licheniformis and its related strains have found extensive applications in diverse industries, agriculture, and medicine. However, the current breeding methods for this strain primarily rely on natural screening and traditional mutagenesis. The limited availability of efficient genetic engineering tools, particularly recombination techniques, has hindered further advancements in its applications. In this study, we conducted a comprehensive investigation to identify and characterize a recombinase, RecT, derived from a Bacillus phage. Remarkably, the recombinase exhibited a 105-fold enhancement in the recombination efficiency of the strain. To facilitate genome editing, we developed a system based on the conditional expression of RecT using a rhamnose-inducible promoter (Prha). The efficacy of this system was evaluated by deleting the amyL gene, which encodes an α-amylase. Our findings revealed that the induction time and concentration of rhamnose, along with the generation time of the strain, significantly influenced the editing efficiency. Optimal conditions for genome editing were determined as follows: the wild-type strain was initially transformed with the genome editing plasmid, followed by cultivation and induction with 1.5% rhamnose for 8 h. Subsequently, the strain was further cultured for an additional 24 h, equivalent to approximately three generations. Consequently, the recombination efficiency reached an impressive 16.67%. This study represents a significant advancement in enhancing the recombination efficiency of B. licheniformis through the utilization of a RecT-based recombination system. Moreover, it provides a highly effective genome editing tool for genetic engineering applications in this strain. Bacillus licheniformis Homologous recombination Recombinase RecT Biotechnology Microbiology Xufan Ma verfasserin aut Cheng Luo verfasserin aut Dongliang Li verfasserin aut Guiyang Shi verfasserin aut Youran Li verfasserin aut In AMB Express SpringerOpen, 2011 13(2023), 1, Seite 15 (DE-627)665672926 (DE-600)2621432-5 21910855 nnns volume:13 year:2023 number:1 pages:15 https://doi.org/10.1186/s13568-023-01589-w kostenfrei https://doaj.org/article/760eeb63251e4ccca3e36bf7af14f284 kostenfrei https://doi.org/10.1186/s13568-023-01589-w kostenfrei https://doaj.org/toc/2191-0855 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2005 GBV_ILN_2009 GBV_ILN_2014 GBV_ILN_2055 GBV_ILN_2111 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 13 2023 1 15
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callnumber-first	T - Technology
author	Fang Xue
spellingShingle	Fang Xue misc TP248.13-248.65 misc QR1-502 misc Bacillus licheniformis misc Homologous recombination misc Recombinase RecT misc Biotechnology misc Microbiology Construction of a bacteriophage-derived recombinase system in Bacillus licheniformis for gene deletion
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topic_title	TP248.13-248.65 QR1-502 Construction of a bacteriophage-derived recombinase system in Bacillus licheniformis for gene deletion Bacillus licheniformis Homologous recombination Recombinase RecT
topic	misc TP248.13-248.65 misc QR1-502 misc Bacillus licheniformis misc Homologous recombination misc Recombinase RecT misc Biotechnology misc Microbiology
topic_unstemmed	misc TP248.13-248.65 misc QR1-502 misc Bacillus licheniformis misc Homologous recombination misc Recombinase RecT misc Biotechnology misc Microbiology
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title	Construction of a bacteriophage-derived recombinase system in Bacillus licheniformis for gene deletion
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title_full	Construction of a bacteriophage-derived recombinase system in Bacillus licheniformis for gene deletion
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title_sort	construction of a bacteriophage-derived recombinase system in bacillus licheniformis for gene deletion
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title_auth	Construction of a bacteriophage-derived recombinase system in Bacillus licheniformis for gene deletion
abstract	Abstract Bacillus licheniformis and its related strains have found extensive applications in diverse industries, agriculture, and medicine. However, the current breeding methods for this strain primarily rely on natural screening and traditional mutagenesis. The limited availability of efficient genetic engineering tools, particularly recombination techniques, has hindered further advancements in its applications. In this study, we conducted a comprehensive investigation to identify and characterize a recombinase, RecT, derived from a Bacillus phage. Remarkably, the recombinase exhibited a 105-fold enhancement in the recombination efficiency of the strain. To facilitate genome editing, we developed a system based on the conditional expression of RecT using a rhamnose-inducible promoter (Prha). The efficacy of this system was evaluated by deleting the amyL gene, which encodes an α-amylase. Our findings revealed that the induction time and concentration of rhamnose, along with the generation time of the strain, significantly influenced the editing efficiency. Optimal conditions for genome editing were determined as follows: the wild-type strain was initially transformed with the genome editing plasmid, followed by cultivation and induction with 1.5% rhamnose for 8 h. Subsequently, the strain was further cultured for an additional 24 h, equivalent to approximately three generations. Consequently, the recombination efficiency reached an impressive 16.67%. This study represents a significant advancement in enhancing the recombination efficiency of B. licheniformis through the utilization of a RecT-based recombination system. Moreover, it provides a highly effective genome editing tool for genetic engineering applications in this strain.
abstractGer	Abstract Bacillus licheniformis and its related strains have found extensive applications in diverse industries, agriculture, and medicine. However, the current breeding methods for this strain primarily rely on natural screening and traditional mutagenesis. The limited availability of efficient genetic engineering tools, particularly recombination techniques, has hindered further advancements in its applications. In this study, we conducted a comprehensive investigation to identify and characterize a recombinase, RecT, derived from a Bacillus phage. Remarkably, the recombinase exhibited a 105-fold enhancement in the recombination efficiency of the strain. To facilitate genome editing, we developed a system based on the conditional expression of RecT using a rhamnose-inducible promoter (Prha). The efficacy of this system was evaluated by deleting the amyL gene, which encodes an α-amylase. Our findings revealed that the induction time and concentration of rhamnose, along with the generation time of the strain, significantly influenced the editing efficiency. Optimal conditions for genome editing were determined as follows: the wild-type strain was initially transformed with the genome editing plasmid, followed by cultivation and induction with 1.5% rhamnose for 8 h. Subsequently, the strain was further cultured for an additional 24 h, equivalent to approximately three generations. Consequently, the recombination efficiency reached an impressive 16.67%. This study represents a significant advancement in enhancing the recombination efficiency of B. licheniformis through the utilization of a RecT-based recombination system. Moreover, it provides a highly effective genome editing tool for genetic engineering applications in this strain.
abstract_unstemmed	Abstract Bacillus licheniformis and its related strains have found extensive applications in diverse industries, agriculture, and medicine. However, the current breeding methods for this strain primarily rely on natural screening and traditional mutagenesis. The limited availability of efficient genetic engineering tools, particularly recombination techniques, has hindered further advancements in its applications. In this study, we conducted a comprehensive investigation to identify and characterize a recombinase, RecT, derived from a Bacillus phage. Remarkably, the recombinase exhibited a 105-fold enhancement in the recombination efficiency of the strain. To facilitate genome editing, we developed a system based on the conditional expression of RecT using a rhamnose-inducible promoter (Prha). The efficacy of this system was evaluated by deleting the amyL gene, which encodes an α-amylase. Our findings revealed that the induction time and concentration of rhamnose, along with the generation time of the strain, significantly influenced the editing efficiency. Optimal conditions for genome editing were determined as follows: the wild-type strain was initially transformed with the genome editing plasmid, followed by cultivation and induction with 1.5% rhamnose for 8 h. Subsequently, the strain was further cultured for an additional 24 h, equivalent to approximately three generations. Consequently, the recombination efficiency reached an impressive 16.67%. This study represents a significant advancement in enhancing the recombination efficiency of B. licheniformis through the utilization of a RecT-based recombination system. Moreover, it provides a highly effective genome editing tool for genetic engineering applications in this strain.
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title_short	Construction of a bacteriophage-derived recombinase system in Bacillus licheniformis for gene deletion
url	https://doi.org/10.1186/s13568-023-01589-w https://doaj.org/article/760eeb63251e4ccca3e36bf7af14f284 https://doaj.org/toc/2191-0855
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Construction of a bacteriophage-derived recombinase system in Bacillus licheniformis for gene deletion

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