Disruption of a DNA fragment that encodes the microneme adhesive repeat domain-containing region of the BBOV_III011730 does not affect the blood stage growth of Babesia bovis in vitro

Bumduuren Tuvshintulga; Azirwan Guswanto; Arifin Budiman Nugraha; Thillaiampalam Sivakumar; Rika Umemiya-Shirafuji; Naoaki Yokoyama

doi:10.1016/j.molbiopara.2023.111576

Disruption of a DNA fragment that encodes the microneme adhesive repeat domain-containing region of the BBOV_III011730 does not affect the blood stage growth of Babesia bovis in vitro

Mol Biochem Parasitol. 2023 Sep:255:111576. doi: 10.1016/j.molbiopara.2023.111576. Epub 2023 Jun 12.

Authors

Bumduuren Tuvshintulga¹, Azirwan Guswanto¹, Arifin Budiman Nugraha¹, Thillaiampalam Sivakumar¹, Rika Umemiya-Shirafuji¹, Naoaki Yokoyama²

Affiliations

¹ National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido, Japan.
² National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido, Japan. Electronic address: yokoyama@obihiro.ac.jp.

PMID: 37315901
DOI: 10.1016/j.molbiopara.2023.111576

Abstract

Babesia bovis, an intraerythrocytic hemoprotozoan parasite, causes the most pathogenic form of bovine babesiosis, negatively impacting the cattle industry. Comprehensive knowledge of B. bovis biology is necessary for developing control methods. In cattle, B. bovis invades the red blood cells (RBCs) and reproduces asexually. Micronemal proteins, which bind to sialic acid of host cells via their microneme adhesive repeat (MAR) domains, are believed to play a key role in host cell invasion by apicomplexan parasites. In this study, we successfully deleted the region encoding MAR domain of the BBOV_III011730 by integrating a fusion gene of enhanced green fluorescent protein-blasticidin-S-deaminase into the genome of B. bovis. The transgenic B. bovis, lacking the MAR domain of the BBOV_III011730, invaded bovine RBCs in vitro and grew at rates similar to the parental line. In conclusion, our study revealed that the MAR domain is non-essential for the intraerythrocytic development of B. bovis in vitro.

Keywords: BBOV_III011730 gene; Babesia bovis; Blood stage; Gene disruption; Growth; Invasion; Microneme adhesive repeat (MAR) domain.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Babesia bovis* / genetics
Babesia bovis* / metabolism
Babesiosis* / parasitology
Cattle
Cattle Diseases* / parasitology
DNA / metabolism
Erythrocytes / parasitology
Microneme

Substances

DNA