稳定表达红色荧光蛋白-木质素脱氨酶融合基因（rfp-bsd），作为卵形巴贝虫 DNA 转染的选择性标记。

IF 1.1 4区农林科学 Q3 VETERINARY SCIENCES

Journal of Veterinary Medical Science Pub Date : 2024-07-02 Epub Date: 2024-05-15 DOI:10.1292/jvms.24-0111

Nada Arayaskul, Masahito Asada, Atefeh Fathi, Nanang R Ariefta, Kota Komatsu, Keisuke Suganuma, Noboru Inoue, Shin-Ichiro Kawazu

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引用次数: 0

摘要

通过电穿孔将红色荧光蛋白（rfp）-blasticidin脱氨酶（bsd）融合基因转染到卵形巴贝虫的质粒DNA中，并在体外培养条件下用15 μg/mL blasticidin S进行筛选。转染后 2 个月，根据 PCR 检测 rfp-bsd 融合基因的存在情况和荧光显微镜下免疫荧光抗体检测融合蛋白的表达情况，筛选并培养出带有表型 DNA 的转染寄生虫，并对其进行进一步分析。据我们所知，这些结果首次证明了在肌动蛋白启动子的控制下，外显子rfp-bsd融合基因作为转染系统的选择性标记在卵形芽孢杆菌中的表达和稳定性。

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Stable expression of red fluorescent protein-blasticidin deaminase fusion gene (rfp-bsd) as a selectable marker for DNA transfection in Babesia ovata.

The red fluorescent protein (rfp)-blasticidin deaminase (bsd) fusion gene was transfected into Babesia ovata by electroporation with the plasmid DNA and selected with 15 μg/mL of blasticidin S under the in vitro culture condition. The transfected parasite with episomal DNA was selected and cultured for further analysis based on the presence of the rfp-bsd fusion gene by PCR and expression of the fusion protein by immunofluorescence antibody test under fluorescence microscopy for 2 months after the transfection. The results are the first, to our knowledge, to demonstrate the expression and stability of the episomal rfp-bsd fusion gene under the control of actin promoter as a selectable marker for the transfection system in B. ovata.

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来源期刊

Journal of Veterinary Medical Science 农林科学-兽医学

CiteScore

2.30

自引率

8.30%

发文量

230

审稿时长

9-18 weeks

期刊介绍： JVMS is a peer-reviewed journal and publishes a variety of papers on veterinary science from basic research to applied science and clinical research. JVMS is published monthly and consists of twelve issues per year. Papers are from the areas of anatomy, physiology, pharmacology, toxicology, pathology, immunology, microbiology, virology, parasitology, internal medicine, surgery, clinical pathology, theriogenology, avian disease, public health, ethology, and laboratory animal science. Although JVMS has played a role in publishing the scientific achievements of Japanese researchers and clinicians for many years, it now also accepts papers submitted from all over the world.