[1]刘卫刚,韩坤煌,谢芳靖,等.大黄鱼免疫球蛋白M重链基因全长cDNA序列的克隆与表达[J].集美大学学报(自然版),2018,23(6):407-415.
 LIU Weigang,HAN Kunhuang,XIE Fangjing,et al.Full­length cDNA Cloning and Expression of LcIgMH Gene from Larimichthys crocea[J].Journal of Jimei University,2018,23(6):407-415.
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大黄鱼免疫球蛋白M重链基因全长cDNA序列的克隆与表达()
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《集美大学学报(自然版)》[ISSN:1007-7405/CN:35-1186/N]

卷:
第23卷
期数:
2018年第6期
页码:
407-415
栏目:
水产、食品与生物技术
出版日期:
2018-11-28

文章信息/Info

Title:
Full­length cDNA Cloning and Expression of LcIgMH Gene from Larimichthys crocea
作者:
刘卫刚1韩坤煌12谢芳靖1邹鹏飞1张子平23王艺磊12
(1.集美大学水产学院,福建 厦门 361021;2.大黄鱼育种国家重点实验室,宁德市富发水产有限公司,福建 宁德 352103;3.福建农林大学动物科学学院,福建 福州 350002)
Author(s):
LIU Weigang1HAN Kunhuang12XIE Fangjing1ZOU Pengfei1ZHANG Ziping23WANG Yilei12
(1.Fisheries College,Jimei University,Xiamen 361021,China;2.State Key Laboratory of Large Yellow Croaker Breeding,Ningde Fufa Fisheries Company Limited,Ningde 352103,China;3.College of Animal Science,Fujian Agriculture and Forestry University,Fuzhou 350002,China)
关键词:
大黄鱼IgMH副溶血弧菌基因表达定量PCR
Keywords:
Larimichthys croceaIgMHVibrio parahaemolyticusgene expressionqRT-PCR
文献标志码:
A
摘要:
本实验室自建的大黄鱼表达标签数据库中获取了大黄鱼免疫球蛋白M重链(LcIgMH)基因片段,并以此设计SMART-RACE引物,最终得到1917 bp的 全长cDNA序列。经分析发现其含6 bp的5′非编码区(5′UTR)、176 bp的3′UTR以及1738 bp的开放阅读框(ORF);预测的ORF共编码584个氨基酸,蛋白质分子量为65.2 ku,等电点为5.84;利用SingalP 4.1 Server发现其5′端含有信号肽(1-19 aa);BlastP Server的结果显示,其与红笛鲷的IgMH序列一致性最高,达到64%;利用IMGT/Domain Gap Align进行重链可变区与恒定区预测,发现含有1个重链可变区与4个重链恒定区,同时,预测还发现可变区含有典型的3个高变区与4个骨架区结构。实时荧光定量PCR结果显示该基因在头肾与脾脏中大量表达,并显著高于其他各组织,同时雌雄鱼LcIgMH在各组织中表达量均无显著性差异,表明LcIgMH表达与性别无关。注射副溶血弧菌进行感染4 d后,脾脏内LcIgMH表达量最高,并且与对照组之间存在显著性差异(P<0.05),但感染后第8天的表达水平已与对照组无显著性差异(P>005),显示LcIgMH与大黄鱼的免疫应答反应相关。
Abstract:
Based on a fragment of the immunoglobulin heavy chain (LcIgMH) gene from Larimichthys crocea expressed sequence tags database of our lab,a full-length LcIgMH cDNA was obtained by the SMART-RACE.The full-length cDNA is 1917 bp,containing a 6 bp 5′untranslated region (5′UTR),a 176 bp 3′UTR,and a 1738 bp open reading frame (ORF) which encodes 584 amino acids.The relative molecular weight of the protein is 65.2 ku,and its isoelectric point is 5.84.A signal peptide (1-19 aa) was found in its N terminal sequence by SingalP4.1 Server.The BlastP result showed that the sequence identity between LcIgMH and Lutjanus sanguineus IgMH was the highest,reaching 64%.A heavy chain variable region and 4 heavy chain constant regions were found by the IMGT/DomainGapAlign Server.The predicted variable region contains typical 3 hypervariable regions and 4 framework regions.The expression of LcIgMH in tissues was detected by quantitative real­time PCR (qRT-PCR).The results show that LcIgMH is extremely highly expressed in the head kidney and spleen,while almost no expression in other tissues.There is no significant difference of LcIgM expression between tissues from male and female,indicating that the expression of LcIgM is not related to gender.After 4 days of infection with Vibrio parahaemolyticus,the expression level of LcIgMH in spleen was significantly increased compared to the control group (P<0.05).However,the expression level of the experimental group and control group had no significant difference at 8 days after injection.

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更新日期/Last Update: 2019-01-18