Liposome fragment-mediated introduction of multiple plasmids into Bacillus subtilis
Transformation of microorganisms by plasmid introduction is one of the central techniques in modern biotechnology. However, applicable transformation methods for simultaneous introduction of multiple plasmids are still limiting. Here, we reported a liposome-mediated method that efficiently introduce...
| Main Authors: | , |
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| Format: | Article |
| Language: | English |
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Elsevier
2019-07-01
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| Series: | Biochemistry and Biophysics Reports |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2405580819300718 |
| _version_ | 1828786957285588992 |
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| author | Kazuki Ohta Norikazu Ichihashi |
| author_facet | Kazuki Ohta Norikazu Ichihashi |
| author_sort | Kazuki Ohta |
| collection | DOAJ |
| description | Transformation of microorganisms by plasmid introduction is one of the central techniques in modern biotechnology. However, applicable transformation methods for simultaneous introduction of multiple plasmids are still limiting. Here, we reported a liposome-mediated method that efficiently introduces multiple plasmids into B. subtilis. In this method, liposomes containing three kinds of plasmids were mixed with B. subtilis protoplasts in the presence of 36% polyethylene glycol (PEG), and the resultant protoplasts were grown in cell wall-regeneration media. We found that the rates of introduction of multiple plasmids were significantly increased in the presence of liposomes. We also found that an intact liposome structure was not required for introduction, and the presence of phosphatidylglycerol (PG) was important for efficient introduction of multiple plasmids. Therefore, the liposome- or liposome fragment-mediated transformation method reported here can advance studies utilizing multiple plasmids. Keywords: Bacillus subtilis, Liposome, Transformation, Multiple plasmids, Protoplast, Polyethyleneglycol |
| first_indexed | 2024-12-12T00:20:52Z |
| format | Article |
| id | doaj.art-97a5cfb151fd444fae308515885eee10 |
| institution | Directory Open Access Journal |
| issn | 2405-5808 |
| language | English |
| last_indexed | 2024-12-12T00:20:52Z |
| publishDate | 2019-07-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Biochemistry and Biophysics Reports |
| spelling | doaj.art-97a5cfb151fd444fae308515885eee102022-12-22T00:44:44ZengElsevierBiochemistry and Biophysics Reports2405-58082019-07-0118Liposome fragment-mediated introduction of multiple plasmids into Bacillus subtilisKazuki Ohta0Norikazu Ichihashi1Graduate School of Frontier Bioscience, Osaka University, 1-5 Yamadaoka, Suita, Osaka, 565-0871, JapanGraduate School of Frontier Bioscience, Osaka University, 1-5 Yamadaoka, Suita, Osaka, 565-0871, Japan; Graduate School of Arts and Science, Komaba Institute for Science, Universal Biology Institute, The University of Tokyo, 3-8-1 Komaba, Meguro-ku, Tokyo, 153-8902, Japan; Corresponding author. Department of Life Science, Graduate School of Arts and Science, The University of Tokyo, 3-8-1 Komaba, Meguro-ku, Tokyo, 153-8902, Japan.Transformation of microorganisms by plasmid introduction is one of the central techniques in modern biotechnology. However, applicable transformation methods for simultaneous introduction of multiple plasmids are still limiting. Here, we reported a liposome-mediated method that efficiently introduces multiple plasmids into B. subtilis. In this method, liposomes containing three kinds of plasmids were mixed with B. subtilis protoplasts in the presence of 36% polyethylene glycol (PEG), and the resultant protoplasts were grown in cell wall-regeneration media. We found that the rates of introduction of multiple plasmids were significantly increased in the presence of liposomes. We also found that an intact liposome structure was not required for introduction, and the presence of phosphatidylglycerol (PG) was important for efficient introduction of multiple plasmids. Therefore, the liposome- or liposome fragment-mediated transformation method reported here can advance studies utilizing multiple plasmids. Keywords: Bacillus subtilis, Liposome, Transformation, Multiple plasmids, Protoplast, Polyethyleneglycolhttp://www.sciencedirect.com/science/article/pii/S2405580819300718 |
| spellingShingle | Kazuki Ohta Norikazu Ichihashi Liposome fragment-mediated introduction of multiple plasmids into Bacillus subtilis Biochemistry and Biophysics Reports |
| title | Liposome fragment-mediated introduction of multiple plasmids into Bacillus subtilis |
| title_full | Liposome fragment-mediated introduction of multiple plasmids into Bacillus subtilis |
| title_fullStr | Liposome fragment-mediated introduction of multiple plasmids into Bacillus subtilis |
| title_full_unstemmed | Liposome fragment-mediated introduction of multiple plasmids into Bacillus subtilis |
| title_short | Liposome fragment-mediated introduction of multiple plasmids into Bacillus subtilis |
| title_sort | liposome fragment mediated introduction of multiple plasmids into bacillus subtilis |
| url | http://www.sciencedirect.com/science/article/pii/S2405580819300718 |
| work_keys_str_mv | AT kazukiohta liposomefragmentmediatedintroductionofmultipleplasmidsintobacillussubtilis AT norikazuichihashi liposomefragmentmediatedintroductionofmultipleplasmidsintobacillussubtilis |