Stable transformation of fluorescent proteins into Nosema bombycis by electroporation.
Parasit Vectors
; 15(1): 141, 2022 Apr 21.
Article
em En
| MEDLINE
| ID: mdl-35449112
ABSTRACT
BACKGROUND:
Microsporidia are a group of intracellular parasitic eukaryotes, serious pathogens that cause widespread infection in humans, vertebrates, and invertebrates. Because microsporidia have a thick spore wall structure, the in vitro transformation, cell culture, and genetic operation technology of microsporidia are far behind that of other parasites.METHODS:
In this study, according to an analysis of the life-cycle of microsporidia, Nosema bombycis, and different electro-transformation conditions, the transduction efficiency of introducing foreign genes into N. bombycis was systematically determined.RESULTS:
We analyzed the direct electro-transformation of foreign genes into germinating N. bombycis using reporters under the regulation of different characteristic promoters. Furthermore, we systematically determined the efficiency of electro-transformation into N. bombycis under different electro-transformation conditions and different developmental stages through an analysis of the whole life-cycle of N. bombycis. These results revealed that foreign genes could be effectively introduced through a perforation voltage of 100 V pulsed for 15 ms during the period of N. bombycis sporeplasm proliferation.CONCLUSIONS:
We present an effective method for electro-transformation of a plasmid encoding a fluorescent protein into N. bombycis, which provides new insight for establishing genetic modifications and potential applications in these intracellular parasites.Palavras-chave
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Bombyx
/
Nosema
Idioma:
En
Ano de publicação:
2022
Tipo de documento:
Article