Expression of an antimicrobial peptide persulcatusin fused with calmodulin in rice cultured cells.

IF 2 3区 生物学 Q2 BIOCHEMICAL RESEARCH METHODS
Gaku Fujita, So Shimoda, Minako Itagaki, Takuto Yahara, Ryuta Tobe, Hiroshi Yoneyama, Yukihiro Ito
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引用次数: 0

Abstract

Antimicrobial proteins and peptides are potential alternatives to antibiotics. Persulcatusin is an antimicrobial peptide found in the taiga tick Ixodes persulcatus. We constructed fusion genes that encode, from the N-terminus to the C-terminus, a signal sequence of rice α-amylase 3D, mouse calmodulin, a target sequence of TEV protease of tobacco etch virus, and persulcatusin with or without a His tag at the N-terminus of the mature fusion protein. These fusion genes were then introduced into rice. Western blot analysis detected persulcatusin fusion proteins in transgenic calli, suspension cells, and their culture medium. Antimicrobial activity against Staphylococcus aureus was detected in the protein extracts prepared from the transgenic callus but not from the non-transgenic wild-type callus, and TEV protease treatment to release persulcatusin from the fusion protein enhanced antimicrobial activity. The growth of the transgenic rice plants was unaffected. Our results indicate that functional persulcatusin can be produced in rice cells. This provides a basis for the mass production of persulcatusin for therapeutic use against bacterial infectious diseases in humans and livestock.

抑菌肽过硫酶与钙调蛋白融合在水稻培养细胞中的表达。
抗菌蛋白和肽是抗生素的潜在替代品。过sulcatusin是在针叶林蜱中发现的一种抗菌肽。我们构建的融合基因从n端到c端分别编码水稻α-淀粉酶3D信号序列、小鼠钙调蛋白、烟草蚀刻病毒TEV蛋白酶靶序列以及成熟融合蛋白n端带或不带His标记的过硫酶。然后将这些融合基因引入水稻。Western blot检测转基因愈伤组织、悬浮细胞及其培养基中过硫凝素融合蛋白。从转基因愈伤组织中提取的蛋白提取物对金黄色葡萄球菌具有抑菌活性,而从非转基因野生型愈伤组织中提取的蛋白提取物则未检测到抑菌活性,并且通过TEV蛋白酶处理使融合蛋白释放过sulcatusin增强了抑菌活性。转基因水稻植株的生长不受影响。我们的研究结果表明,功能性过硫酶可以在水稻细胞中产生。这为大规模生产用于治疗人类和牲畜细菌传染病的过硫酶提供了基础。
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来源期刊
Transgenic Research
Transgenic Research 生物-生化研究方法
CiteScore
5.40
自引率
0.00%
发文量
38
审稿时长
4-8 weeks
期刊介绍: Transgenic Research focusses on transgenic and genome edited higher organisms. Manuscripts emphasizing biotechnological applications are strongly encouraged. Intellectual property, ethical issues, societal impact and regulatory aspects also fall within the scope of the journal. Transgenic Research aims to bridge the gap between fundamental and applied science in molecular biology and biotechnology for the plant and animal academic and associated industry communities. Transgenic Research publishes -Original Papers -Reviews: Should critically summarize the current state-of-the-art of the subject in a dispassionate way. Authors are requested to contact a Board Member before submission. Reviews should not be descriptive; rather they should present the most up-to-date information on the subject in a dispassionate and critical way. Perspective Reviews which can address new or controversial aspects are encouraged. -Brief Communications: Should report significant developments in methodology and experimental transgenic higher organisms
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