Tracing endogenous proteins in living cells through electrotransfer of mRNA encoding chromobodies
Biotechnol J. 2024 Feb;19(2):e2300548. doi: 10.1002/biot.202300548.ABSTRACTChromobodies made of nanobodies fused to fluorescent proteins are powerful tools for targeting and tracing intracellular proteins in living cells. Typically, this is achieved by transfecting plasmids encoding the chromobodies. However, an excess of unbound chromobody relative to the endogenous antigen can result in high background fluorescence in live cell imaging. Here, we overcome this problem by using mRNA encoding chromobodies. Our approach allows one to precisely control the amount of chromobody expressed inside the cell by adjusting the amount...
Source: Biotechnology Journal - February 26, 2024 Category: Biotechnology Authors: Th éo Juncker Ludovic Richert Murielle Masson Guy Zuber Bruno Chatton Mariel Donzeau Source Type: research

Liamocins overproduction via the two-pH stage fermentation and anti-Aspergillus flavus activity of Massoia lactone
Biotechnol J. 2024 Feb;19(2):e2300675. doi: 10.1002/biot.202300675.ABSTRACTAureobasidium melanogenum was found to be grown the best at the constant pH 7.0 and to produce the highest amount of liamocins at the constant pH 3.0. Therefore, the wild type strain A. melanogenum 9-1 and the engineered strain V33 constructed in the laboratory were grown at the constant pH 7.0 for 48 h, then, they were continued to be cultivated at the constant pH 3.0. Under such conditions, A. melanogenum 9-1 produced 36.51 ± 0.55 g L-1 of liamocin and its cell mass was 27.43 ± 0.63 and 6.00 ± 0.11 g L-1 of glucose was left in the finished medi...
Source: Biotechnology Journal - February 26, 2024 Category: Biotechnology Authors: Peng Wang Mei Zhang Shou-Feng Zhao Zhao-Rui Zhang Guang-Lei Liu Zhe Chi Zhen-Ming Chi Source Type: research

Tracing endogenous proteins in living cells through electrotransfer of mRNA encoding chromobodies
Biotechnol J. 2024 Feb;19(2):e2300548. doi: 10.1002/biot.202300548.ABSTRACTChromobodies made of nanobodies fused to fluorescent proteins are powerful tools for targeting and tracing intracellular proteins in living cells. Typically, this is achieved by transfecting plasmids encoding the chromobodies. However, an excess of unbound chromobody relative to the endogenous antigen can result in high background fluorescence in live cell imaging. Here, we overcome this problem by using mRNA encoding chromobodies. Our approach allows one to precisely control the amount of chromobody expressed inside the cell by adjusting the amount...
Source: Biotechnology Journal - February 26, 2024 Category: Biotechnology Authors: Th éo Juncker Ludovic Richert Murielle Masson Guy Zuber Bruno Chatton Mariel Donzeau Source Type: research

Liamocins overproduction via the two-pH stage fermentation and anti-Aspergillus flavus activity of Massoia lactone
Biotechnol J. 2024 Feb;19(2):e2300675. doi: 10.1002/biot.202300675.ABSTRACTAureobasidium melanogenum was found to be grown the best at the constant pH 7.0 and to produce the highest amount of liamocins at the constant pH 3.0. Therefore, the wild type strain A. melanogenum 9-1 and the engineered strain V33 constructed in the laboratory were grown at the constant pH 7.0 for 48 h, then, they were continued to be cultivated at the constant pH 3.0. Under such conditions, A. melanogenum 9-1 produced 36.51 ± 0.55 g L-1 of liamocin and its cell mass was 27.43 ± 0.63 and 6.00 ± 0.11 g L-1 of glucose was left in the finished medi...
Source: Biotechnology Journal - February 26, 2024 Category: Biotechnology Authors: Peng Wang Mei Zhang Shou-Feng Zhao Zhao-Rui Zhang Guang-Lei Liu Zhe Chi Zhen-Ming Chi Source Type: research

Tracing endogenous proteins in living cells through electrotransfer of mRNA encoding chromobodies
Biotechnol J. 2024 Feb;19(2):e2300548. doi: 10.1002/biot.202300548.ABSTRACTChromobodies made of nanobodies fused to fluorescent proteins are powerful tools for targeting and tracing intracellular proteins in living cells. Typically, this is achieved by transfecting plasmids encoding the chromobodies. However, an excess of unbound chromobody relative to the endogenous antigen can result in high background fluorescence in live cell imaging. Here, we overcome this problem by using mRNA encoding chromobodies. Our approach allows one to precisely control the amount of chromobody expressed inside the cell by adjusting the amount...
Source: Biotechnology Journal - February 26, 2024 Category: Biotechnology Authors: Th éo Juncker Ludovic Richert Murielle Masson Guy Zuber Bruno Chatton Mariel Donzeau Source Type: research

Liamocins overproduction via the two-pH stage fermentation and anti-Aspergillus flavus activity of Massoia lactone
Biotechnol J. 2024 Feb;19(2):e2300675. doi: 10.1002/biot.202300675.ABSTRACTAureobasidium melanogenum was found to be grown the best at the constant pH 7.0 and to produce the highest amount of liamocins at the constant pH 3.0. Therefore, the wild type strain A. melanogenum 9-1 and the engineered strain V33 constructed in the laboratory were grown at the constant pH 7.0 for 48 h, then, they were continued to be cultivated at the constant pH 3.0. Under such conditions, A. melanogenum 9-1 produced 36.51 ± 0.55 g L-1 of liamocin and its cell mass was 27.43 ± 0.63 and 6.00 ± 0.11 g L-1 of glucose was left in the finished medi...
Source: Biotechnology Journal - February 26, 2024 Category: Biotechnology Authors: Peng Wang Mei Zhang Shou-Feng Zhao Zhao-Rui Zhang Guang-Lei Liu Zhe Chi Zhen-Ming Chi Source Type: research

Tracing endogenous proteins in living cells through electrotransfer of mRNA encoding chromobodies
Biotechnol J. 2024 Feb;19(2):e2300548. doi: 10.1002/biot.202300548.ABSTRACTChromobodies made of nanobodies fused to fluorescent proteins are powerful tools for targeting and tracing intracellular proteins in living cells. Typically, this is achieved by transfecting plasmids encoding the chromobodies. However, an excess of unbound chromobody relative to the endogenous antigen can result in high background fluorescence in live cell imaging. Here, we overcome this problem by using mRNA encoding chromobodies. Our approach allows one to precisely control the amount of chromobody expressed inside the cell by adjusting the amount...
Source: Biotechnology Journal - February 26, 2024 Category: Biotechnology Authors: Th éo Juncker Ludovic Richert Murielle Masson Guy Zuber Bruno Chatton Mariel Donzeau Source Type: research

Liamocins overproduction via the two-pH stage fermentation and anti-Aspergillus flavus activity of Massoia lactone
Biotechnol J. 2024 Feb;19(2):e2300675. doi: 10.1002/biot.202300675.ABSTRACTAureobasidium melanogenum was found to be grown the best at the constant pH 7.0 and to produce the highest amount of liamocins at the constant pH 3.0. Therefore, the wild type strain A. melanogenum 9-1 and the engineered strain V33 constructed in the laboratory were grown at the constant pH 7.0 for 48 h, then, they were continued to be cultivated at the constant pH 3.0. Under such conditions, A. melanogenum 9-1 produced 36.51 ± 0.55 g L-1 of liamocin and its cell mass was 27.43 ± 0.63 and 6.00 ± 0.11 g L-1 of glucose was left in the finished medi...
Source: Biotechnology Journal - February 26, 2024 Category: Biotechnology Authors: Peng Wang Mei Zhang Shou-Feng Zhao Zhao-Rui Zhang Guang-Lei Liu Zhe Chi Zhen-Ming Chi Source Type: research

Tracing endogenous proteins in living cells through electrotransfer of mRNA encoding chromobodies
Biotechnol J. 2024 Feb;19(2):e2300548. doi: 10.1002/biot.202300548.ABSTRACTChromobodies made of nanobodies fused to fluorescent proteins are powerful tools for targeting and tracing intracellular proteins in living cells. Typically, this is achieved by transfecting plasmids encoding the chromobodies. However, an excess of unbound chromobody relative to the endogenous antigen can result in high background fluorescence in live cell imaging. Here, we overcome this problem by using mRNA encoding chromobodies. Our approach allows one to precisely control the amount of chromobody expressed inside the cell by adjusting the amount...
Source: Biotechnology Journal - February 26, 2024 Category: Biotechnology Authors: Th éo Juncker Ludovic Richert Murielle Masson Guy Zuber Bruno Chatton Mariel Donzeau Source Type: research

Liamocins overproduction via the two-pH stage fermentation and anti-Aspergillus flavus activity of Massoia lactone
Biotechnol J. 2024 Feb;19(2):e2300675. doi: 10.1002/biot.202300675.ABSTRACTAureobasidium melanogenum was found to be grown the best at the constant pH 7.0 and to produce the highest amount of liamocins at the constant pH 3.0. Therefore, the wild type strain A. melanogenum 9-1 and the engineered strain V33 constructed in the laboratory were grown at the constant pH 7.0 for 48 h, then, they were continued to be cultivated at the constant pH 3.0. Under such conditions, A. melanogenum 9-1 produced 36.51 ± 0.55 g L-1 of liamocin and its cell mass was 27.43 ± 0.63 and 6.00 ± 0.11 g L-1 of glucose was left in the finished medi...
Source: Biotechnology Journal - February 26, 2024 Category: Biotechnology Authors: Peng Wang Mei Zhang Shou-Feng Zhao Zhao-Rui Zhang Guang-Lei Liu Zhe Chi Zhen-Ming Chi Source Type: research

Tracing endogenous proteins in living cells through electrotransfer of mRNA encoding chromobodies
Biotechnol J. 2024 Feb;19(2):e2300548. doi: 10.1002/biot.202300548.ABSTRACTChromobodies made of nanobodies fused to fluorescent proteins are powerful tools for targeting and tracing intracellular proteins in living cells. Typically, this is achieved by transfecting plasmids encoding the chromobodies. However, an excess of unbound chromobody relative to the endogenous antigen can result in high background fluorescence in live cell imaging. Here, we overcome this problem by using mRNA encoding chromobodies. Our approach allows one to precisely control the amount of chromobody expressed inside the cell by adjusting the amount...
Source: Biotechnology Journal - February 26, 2024 Category: Biotechnology Authors: Th éo Juncker Ludovic Richert Murielle Masson Guy Zuber Bruno Chatton Mariel Donzeau Source Type: research

Liamocins overproduction via the two-pH stage fermentation and anti-Aspergillus flavus activity of Massoia lactone
Biotechnol J. 2024 Feb;19(2):e2300675. doi: 10.1002/biot.202300675.ABSTRACTAureobasidium melanogenum was found to be grown the best at the constant pH 7.0 and to produce the highest amount of liamocins at the constant pH 3.0. Therefore, the wild type strain A. melanogenum 9-1 and the engineered strain V33 constructed in the laboratory were grown at the constant pH 7.0 for 48 h, then, they were continued to be cultivated at the constant pH 3.0. Under such conditions, A. melanogenum 9-1 produced 36.51 ± 0.55 g L-1 of liamocin and its cell mass was 27.43 ± 0.63 and 6.00 ± 0.11 g L-1 of glucose was left in the finished medi...
Source: Biotechnology Journal - February 26, 2024 Category: Biotechnology Authors: Peng Wang Mei Zhang Shou-Feng Zhao Zhao-Rui Zhang Guang-Lei Liu Zhe Chi Zhen-Ming Chi Source Type: research

Tracing endogenous proteins in living cells through electrotransfer of mRNA encoding chromobodies
Biotechnol J. 2024 Feb;19(2):e2300548. doi: 10.1002/biot.202300548.ABSTRACTChromobodies made of nanobodies fused to fluorescent proteins are powerful tools for targeting and tracing intracellular proteins in living cells. Typically, this is achieved by transfecting plasmids encoding the chromobodies. However, an excess of unbound chromobody relative to the endogenous antigen can result in high background fluorescence in live cell imaging. Here, we overcome this problem by using mRNA encoding chromobodies. Our approach allows one to precisely control the amount of chromobody expressed inside the cell by adjusting the amount...
Source: Biotechnology Journal - February 26, 2024 Category: Biotechnology Authors: Th éo Juncker Ludovic Richert Murielle Masson Guy Zuber Bruno Chatton Mariel Donzeau Source Type: research

Liamocins overproduction via the two-pH stage fermentation and anti-Aspergillus flavus activity of Massoia lactone
Biotechnol J. 2024 Feb;19(2):e2300675. doi: 10.1002/biot.202300675.ABSTRACTAureobasidium melanogenum was found to be grown the best at the constant pH 7.0 and to produce the highest amount of liamocins at the constant pH 3.0. Therefore, the wild type strain A. melanogenum 9-1 and the engineered strain V33 constructed in the laboratory were grown at the constant pH 7.0 for 48 h, then, they were continued to be cultivated at the constant pH 3.0. Under such conditions, A. melanogenum 9-1 produced 36.51 ± 0.55 g L-1 of liamocin and its cell mass was 27.43 ± 0.63 and 6.00 ± 0.11 g L-1 of glucose was left in the finished medi...
Source: Biotechnology Journal - February 26, 2024 Category: Biotechnology Authors: Peng Wang Mei Zhang Shou-Feng Zhao Zhao-Rui Zhang Guang-Lei Liu Zhe Chi Zhen-Ming Chi Source Type: research

Tracing endogenous proteins in living cells through electrotransfer of mRNA encoding chromobodies
Biotechnol J. 2024 Feb;19(2):e2300548. doi: 10.1002/biot.202300548.ABSTRACTChromobodies made of nanobodies fused to fluorescent proteins are powerful tools for targeting and tracing intracellular proteins in living cells. Typically, this is achieved by transfecting plasmids encoding the chromobodies. However, an excess of unbound chromobody relative to the endogenous antigen can result in high background fluorescence in live cell imaging. Here, we overcome this problem by using mRNA encoding chromobodies. Our approach allows one to precisely control the amount of chromobody expressed inside the cell by adjusting the amount...
Source: Biotechnology Journal - February 26, 2024 Category: Biotechnology Authors: Th éo Juncker Ludovic Richert Murielle Masson Guy Zuber Bruno Chatton Mariel Donzeau Source Type: research