文献类型：期刊文献

中文题名：凡纳滨对虾ARMC8基因的克隆及表达

英文题名：Cloning and expression profile analysis of ARMC8 gene from Litopenaeus vannamei

作者：黄金凤[1];杨奇慧[1,2];董晓慧[1,2];迟淑艳[1,2];刘泓宇[1,2];谭北平[1,2];章双[1,2]

机构：[1]广东海洋大学水产学院,广东湛江524088;[2]南海生物资源开发与利用协同创新中心,广东珠海519082

年份：2017

卷号：41

期号：2

起止页码：171

中文期刊名：水产学报

外文期刊名：Journal of Fisheries of China

收录：CSTPCD、、北大核心2014、Scopus、CSCD2017_2018、北大核心、CSCD

基金：广东省自然科学基金-博士启动项目(2014A030310184);广东省自然科学基金-自由申请项目(2016A030313757);广东普通高校青年创新人才项目(2014KQNCX079);广东海洋大学优秀青年骨干教师特别资助计划(HDYQ2015004)~~

语种：中文

中文关键词：凡纳滨对虾;ARMC8基因;基因克隆;组织表达;WSSV感染;副溶血弧菌

外文关键词：Litopenaeus vannamei; armadillo repeat-containing protein 8; gene cloning; tissue expression; WSSV infection; Vibrio parahaemolyticus

中文摘要：为了研究含Armadillo重复蛋白8在凡纳滨对虾抗病感染过程中所起的免疫作用,本实验采用RACE-PCR技术首次克隆得到凡纳滨对虾ARMC8基因(LvARMC8,Gen Bank注册号:KX058562)的c DNA序列全长,并利用在线软件进行生物信息学分析;采用实时荧光定量PCR(Rt-PCR)技术对LvARMC8基因在凡纳滨对虾不同组织及感染白斑综合征病毒和副溶血弧菌过程中的表达变化特征进行分析。结果显示,LvARMC8基因全长为2917bp,其中开放阅读框长2046 bp,编码681个氨基酸,5′非编码区长49 bp,3′非编码区长822 bp。预测分析显示LvARMC8编码的蛋白质含有6个ARM结构域。同源性分析发现,LvARMC8基因与内华达古白蚁ARMC8基因的相似度最高,为71%。系统进化分析结果显示,LvARMC8和多种无脊椎动物ARMC8聚为一支,其中与昆虫类动物赤拟谷盗、致倦库蚊和柑橘凤蝶的亲缘关系最近。Rt-PCR分析发现,LvARMC8基因在凡纳滨对虾多个组织中均能检测出,在表皮中表达量最高,眼柄中表达量最低。WSSV感染后12 h,LvARMC8基因在凡纳滨对虾血液中的表达量显著降低,但48 h后显著升高,72 h达到最高水平。除副溶血弧菌感染后24 h外的时间点,LvARMC8基因在凡纳滨对虾血液中的表达量均显著升高。研究表明,LvARMC8基因可能参与凡纳滨对虾抗病免疫应答途径。

外文摘要：The purpose is to study the potential role of Litopenaeus vannamei armadillo repeat-containing protein 8（ARMC8） in the immune response triggered by the virus and bacterial pathogens. Full-length c DNA sequence of ARMC8 gene from L. vannamei（named LvARMC8, Gen Bank Accession Number： KX058562） was first cloned using RACE method. The full-length c DNA sequence of LvARMC8 was 2917 bp, which contains a 50 bp 5＇UTR,822 bp 3＇UTR and 2046 bp open reading frame（ORF） that encoded 681 amino acid residues. SMART analysis results showed that LvARMC8 contains six armadillo repeat（ARM） domains. Multiple alignment analysis shows that LvARMC8 shared 71% amino acid identity with Zootermopsis nevadensis, which is the highest. Phylogenetic analysis showed that LvARMC8 was clustered together of invertebrates groups and most closely related to Tribolium castaneum ARMC8, Culex quinquefasciatus ARMC8 and Papilio xuthus ARMC8. RT-PCR analysis showed that LvARMC8 was constitutively expressed in all the examined tissues with the highest expression in epithelium and lowest expression in eyestalk. Upon WSSV challenge, the expression of LvARMC8 was significantly down-regulated at 12 hpi（hours past infection） but significantly up-regulated starting at 48 hpi,reaching the peak at 72 hpi. By V. parahaemolyticus challenging, the level of LvARMC8 expression were markedly increased at all the detected time points except 24 hpi. The result suggests that LvARMC8 might take part in the innate immune response of L. vannamei triggered by pathogens.