Improvement of an in vitro drug selection method for generating transgenic Plasmodium berghei parasites
Abstract Background Reverse genetics approaches have become powerful tools to dissect the biology of malaria parasites. In a previous study, development of an in vitro drug selection method for generating transgenic parasite of Plasmodium berghei was reported. Using this method, two novel and indepe...
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ftdoajarticles:oai:doaj.org/article:b23b7b8dde04403093dd186adc7b98c9 2023-05-15T15:12:26+02:00 Improvement of an in vitro drug selection method for generating transgenic Plasmodium berghei parasites Akira Soga Takahiro Shirozu Mami Ko-ketsu Shinya Fukumoto 2019-06-01T00:00:00Z https://doi.org/10.1186/s12936-019-2851-6 https://doaj.org/article/b23b7b8dde04403093dd186adc7b98c9 EN eng BMC http://link.springer.com/article/10.1186/s12936-019-2851-6 https://doaj.org/toc/1475-2875 doi:10.1186/s12936-019-2851-6 1475-2875 https://doaj.org/article/b23b7b8dde04403093dd186adc7b98c9 Malaria Journal, Vol 18, Iss 1, Pp 1-9 (2019) Plasmodium berghei Transgenic parasite Drug selection Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 article 2019 ftdoajarticles https://doi.org/10.1186/s12936-019-2851-6 2022-12-31T01:04:34Z Abstract Background Reverse genetics approaches have become powerful tools to dissect the biology of malaria parasites. In a previous study, development of an in vitro drug selection method for generating transgenic parasite of Plasmodium berghei was reported. Using this method, two novel and independent selection markers using the P. berghei heat shock protein 70 promoter was previously established. While the approach permits the easy and flexible genetic manipulation of P. berghei, shortcomings include a low variety in promoter options to drive marker gene expression and increased complexity of the selection procedure. In this study, addressing these issues was attempted. Methods To secure a variety of promoters, the use of a P. berghei elongation factor-1α promoter for marker gene expression was attempted. To simplify the procedure of in vitro selection, the establishment of a two cell-cycle culture method and its application for drug selection were attempted. Results The P. berghei elongation factor-1α (pbef-1α) promoter, which is commonly used to drive marker gene expression, was successfully applied as an alternative promoter model for marker gene expression, using the parasite’s codon-optimized marker sequence. To simplify the in vitro selection method, a two cell-cycle culture method in which the merozoite was released by filtration of the culture containing matured schizont-infected erythrocytes was also developed and successfully applied for drug selection. Conclusion The pbef-1α promoter was successfully applied in an in vitro selection system. The in vitro selection procedure also could be simplified for practical use using a two cell-cycle culture method. These improvements provide a more versatile platform for the genetic manipulation of P. berghei. Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic Malaria Journal 18 1 |
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Directory of Open Access Journals: DOAJ Articles |
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English |
topic |
Plasmodium berghei Transgenic parasite Drug selection Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 |
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Plasmodium berghei Transgenic parasite Drug selection Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 Akira Soga Takahiro Shirozu Mami Ko-ketsu Shinya Fukumoto Improvement of an in vitro drug selection method for generating transgenic Plasmodium berghei parasites |
topic_facet |
Plasmodium berghei Transgenic parasite Drug selection Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 |
description |
Abstract Background Reverse genetics approaches have become powerful tools to dissect the biology of malaria parasites. In a previous study, development of an in vitro drug selection method for generating transgenic parasite of Plasmodium berghei was reported. Using this method, two novel and independent selection markers using the P. berghei heat shock protein 70 promoter was previously established. While the approach permits the easy and flexible genetic manipulation of P. berghei, shortcomings include a low variety in promoter options to drive marker gene expression and increased complexity of the selection procedure. In this study, addressing these issues was attempted. Methods To secure a variety of promoters, the use of a P. berghei elongation factor-1α promoter for marker gene expression was attempted. To simplify the procedure of in vitro selection, the establishment of a two cell-cycle culture method and its application for drug selection were attempted. Results The P. berghei elongation factor-1α (pbef-1α) promoter, which is commonly used to drive marker gene expression, was successfully applied as an alternative promoter model for marker gene expression, using the parasite’s codon-optimized marker sequence. To simplify the in vitro selection method, a two cell-cycle culture method in which the merozoite was released by filtration of the culture containing matured schizont-infected erythrocytes was also developed and successfully applied for drug selection. Conclusion The pbef-1α promoter was successfully applied in an in vitro selection system. The in vitro selection procedure also could be simplified for practical use using a two cell-cycle culture method. These improvements provide a more versatile platform for the genetic manipulation of P. berghei. |
format |
Article in Journal/Newspaper |
author |
Akira Soga Takahiro Shirozu Mami Ko-ketsu Shinya Fukumoto |
author_facet |
Akira Soga Takahiro Shirozu Mami Ko-ketsu Shinya Fukumoto |
author_sort |
Akira Soga |
title |
Improvement of an in vitro drug selection method for generating transgenic Plasmodium berghei parasites |
title_short |
Improvement of an in vitro drug selection method for generating transgenic Plasmodium berghei parasites |
title_full |
Improvement of an in vitro drug selection method for generating transgenic Plasmodium berghei parasites |
title_fullStr |
Improvement of an in vitro drug selection method for generating transgenic Plasmodium berghei parasites |
title_full_unstemmed |
Improvement of an in vitro drug selection method for generating transgenic Plasmodium berghei parasites |
title_sort |
improvement of an in vitro drug selection method for generating transgenic plasmodium berghei parasites |
publisher |
BMC |
publishDate |
2019 |
url |
https://doi.org/10.1186/s12936-019-2851-6 https://doaj.org/article/b23b7b8dde04403093dd186adc7b98c9 |
geographic |
Arctic |
geographic_facet |
Arctic |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
Malaria Journal, Vol 18, Iss 1, Pp 1-9 (2019) |
op_relation |
http://link.springer.com/article/10.1186/s12936-019-2851-6 https://doaj.org/toc/1475-2875 doi:10.1186/s12936-019-2851-6 1475-2875 https://doaj.org/article/b23b7b8dde04403093dd186adc7b98c9 |
op_doi |
https://doi.org/10.1186/s12936-019-2851-6 |
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Malaria Journal |
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18 |
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1 |
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1766343114441621504 |