刘恩慧,黄天晴,谷伟,王炳谦,张艳萍,李春雨,徐革锋

• 1:中国水产科学研究院黑龙江水产研究所农业农村部淡水水产生物技术与遗传育种重点实验室
• 2:甘肃省水产研究所
• 3:新疆天蕴有机农业有限公司

摘要(Abstract)：

为探究乌苏里白鲑Coregonus ussuriensis Berg肝脏表达抗菌肽Leap-2基因的分子结构和表达特征，采用PCR技术从乌苏里白鲑(体质量为30.0 g±1.2 g)肝脏组织中扩增Leap-2基因的开放阅读框(ORF),使用生物信息学软件对其编码的氨基酸序列进行分析，通过荧光定量PCR(qRT-PCR)检测其在健康组织及嗜水气单胞菌Aeromonas hydrophila、杀鲑气单胞菌Aeromonas salmonicida和金黄色葡萄球菌Staphylococcus aureus感染后的组织表达变化。结果表明：采用PCR方法从乌苏里白鲑肝脏中克隆得到Leap-2基因的ORF区(GenBank:MZ170050),片段长度为276 bp,可编码91个氨基酸，其成熟肽区域包含4个高度保守的半胱氨酸残基；系统进化分析显示，乌苏里白鲑的Leap-2氨基酸序列与银鲑Oncorhynchus kisutch的亲缘关系较近，且具有较高的同源性，一致性为96.7%;乌苏里白鲑肝脏中的Leap-2基因表达量最高，其次为肠和肾，肌肉中的基因表达量最低；3种细菌感染导致Leap-2在各组织中的表达均有所上调，Western blot检测也显示，Leap-2蛋白水平在24 h内表达上调。研究表明，Leap-2基因参与了乌苏里白鲑应对外源干扰的免疫反应，本研究结果可为解析乌苏里白鲑应对病原体的免疫应答机制提供科学参考。

关键词(KeyWords)： 乌苏里白鲑;抗菌肽;Leap-2基因;嗜水气单胞菌;杀鲑气单胞菌;金黄色葡萄球菌;荧光定量

Abstract:

Keywords:

基金项目(Foundation): 中央级公益性科研院所基本科研业务费专项(HSY202103M);; 财政部和农业农村部：国家现代农业产业技术体系资助(CARS-46);; 中国水产科学研究院科研业务费项目(2020TD32);; 农业财政专项“东北地区重点水域渔业资源与环境调查”

作者(Author): 刘恩慧,黄天晴,谷伟,王炳谦,张艳萍,李春雨,徐革锋

DOI: 10.16535/j.cnki.dlhyxb.2021-163

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