Two Foreign Antimicrobial Peptides Expressed in the Chloroplast of <i>Porphyridium purpureum</i> Possessed Antibacterial Properties
To solve the problem of antibiotic abuse in aquaculture and to utilize the application potential of antimicrobial peptides (AMPs), a chloroplast transformation system of <i>Porphyridium purpureum</i> was successfully constructed for effectively expressing two exogenous AMPs. The endogeno...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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MDPI AG
2022-07-01
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| Series: | Marine Drugs |
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| Online Access: | https://www.mdpi.com/1660-3397/20/8/484 |
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| author | Subing Han Jialin Zhao Ping Liu Kang Wang Song Qin Zhenjun Zhao Yulin Cui |
| author_facet | Subing Han Jialin Zhao Ping Liu Kang Wang Song Qin Zhenjun Zhao Yulin Cui |
| author_sort | Subing Han |
| collection | DOAJ |
| description | To solve the problem of antibiotic abuse in aquaculture and to utilize the application potential of antimicrobial peptides (AMPs), a chloroplast transformation system of <i>Porphyridium purpureum</i> was successfully constructed for effectively expressing two exogenous AMPs. The endogenous fragments of 16S rDNA/<i>trnA</i>-23S rDNA were used as flanking fragments for the homologous recombination in the chloroplast genome. Two AMPs encoded by the transformation vector were controlled by the native promoter <i>psbB</i> in a polycistron. The plasmids were transferred into <i>P. purpureum</i> via particle bombardment and the transformation vectors were screened using phosphinothricin (<i>bar</i>), a dominant selection marker under the control of the <i>psbA</i> promoter. Subsequently, in the positive transformed colonies, the exogenous fragments were found to be inserted in the flanking fragments directionally as expected and two foreign AMPs were successfully obtained. Finally, two exogenous peptides with antibacterial properties were obtained from the transformed strain. The two AMPs expressed by the transformed strain were shown to have similar inhibitory effects to antibiotics by inhibition tests. This suggested that AMPs can be introduced into aquaculture using baited microalgae, providing new ideas and ways to solve a series of aquaculture diseases caused by bacteria. |
| first_indexed | 2024-03-09T13:04:14Z |
| format | Article |
| id | doaj.art-ea7cc83ad2224b4587f79b0be74f3d7f |
| institution | Directory Open Access Journal |
| issn | 1660-3397 |
| language | English |
| last_indexed | 2024-03-09T13:04:14Z |
| publishDate | 2022-07-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Marine Drugs |
| spelling | doaj.art-ea7cc83ad2224b4587f79b0be74f3d7f2023-11-30T21:51:41ZengMDPI AGMarine Drugs1660-33972022-07-0120848410.3390/md20080484Two Foreign Antimicrobial Peptides Expressed in the Chloroplast of <i>Porphyridium purpureum</i> Possessed Antibacterial PropertiesSubing Han0Jialin Zhao1Ping Liu2Kang Wang3Song Qin4Zhenjun Zhao5Yulin Cui6College of Life Sciences, Yantai University, Yantai 264000, ChinaYantai Marine Economic Research Institute, Yantai 264003, ChinaCollege of Life Sciences, Yantai University, Yantai 264000, ChinaKey Laboratory of Coastal Biology and Biological Resource Utilization, Yantai Institute of Coastal Zone Research, Chinese Academy of Sciences, Yantai 264003, ChinaKey Laboratory of Coastal Biology and Biological Resource Utilization, Yantai Institute of Coastal Zone Research, Chinese Academy of Sciences, Yantai 264003, ChinaCollege of Life Sciences, Yantai University, Yantai 264000, ChinaKey Laboratory of Coastal Biology and Biological Resource Utilization, Yantai Institute of Coastal Zone Research, Chinese Academy of Sciences, Yantai 264003, ChinaTo solve the problem of antibiotic abuse in aquaculture and to utilize the application potential of antimicrobial peptides (AMPs), a chloroplast transformation system of <i>Porphyridium purpureum</i> was successfully constructed for effectively expressing two exogenous AMPs. The endogenous fragments of 16S rDNA/<i>trnA</i>-23S rDNA were used as flanking fragments for the homologous recombination in the chloroplast genome. Two AMPs encoded by the transformation vector were controlled by the native promoter <i>psbB</i> in a polycistron. The plasmids were transferred into <i>P. purpureum</i> via particle bombardment and the transformation vectors were screened using phosphinothricin (<i>bar</i>), a dominant selection marker under the control of the <i>psbA</i> promoter. Subsequently, in the positive transformed colonies, the exogenous fragments were found to be inserted in the flanking fragments directionally as expected and two foreign AMPs were successfully obtained. Finally, two exogenous peptides with antibacterial properties were obtained from the transformed strain. The two AMPs expressed by the transformed strain were shown to have similar inhibitory effects to antibiotics by inhibition tests. This suggested that AMPs can be introduced into aquaculture using baited microalgae, providing new ideas and ways to solve a series of aquaculture diseases caused by bacteria.https://www.mdpi.com/1660-3397/20/8/484<i>Porphyridium purpureum</i>chloroplast transformation systemmicroparticle bombardmentantimicrobial peptidesNZ2114Piscidin-4 |
| spellingShingle | Subing Han Jialin Zhao Ping Liu Kang Wang Song Qin Zhenjun Zhao Yulin Cui Two Foreign Antimicrobial Peptides Expressed in the Chloroplast of <i>Porphyridium purpureum</i> Possessed Antibacterial Properties Marine Drugs <i>Porphyridium purpureum</i> chloroplast transformation system microparticle bombardment antimicrobial peptides NZ2114 Piscidin-4 |
| title | Two Foreign Antimicrobial Peptides Expressed in the Chloroplast of <i>Porphyridium purpureum</i> Possessed Antibacterial Properties |
| title_full | Two Foreign Antimicrobial Peptides Expressed in the Chloroplast of <i>Porphyridium purpureum</i> Possessed Antibacterial Properties |
| title_fullStr | Two Foreign Antimicrobial Peptides Expressed in the Chloroplast of <i>Porphyridium purpureum</i> Possessed Antibacterial Properties |
| title_full_unstemmed | Two Foreign Antimicrobial Peptides Expressed in the Chloroplast of <i>Porphyridium purpureum</i> Possessed Antibacterial Properties |
| title_short | Two Foreign Antimicrobial Peptides Expressed in the Chloroplast of <i>Porphyridium purpureum</i> Possessed Antibacterial Properties |
| title_sort | two foreign antimicrobial peptides expressed in the chloroplast of i porphyridium purpureum i possessed antibacterial properties |
| topic | <i>Porphyridium purpureum</i> chloroplast transformation system microparticle bombardment antimicrobial peptides NZ2114 Piscidin-4 |
| url | https://www.mdpi.com/1660-3397/20/8/484 |
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