半滑舌鳎黏蛋白MUC5AC基因的克隆与结构预测

doi:10.19640/j.cnki.jtau.2019.02.012

天津农学院学报 ›› 2019, Vol. 26 ›› Issue (2): 51-56.doi: 10.19640/j.cnki.jtau.2019.02.012

半滑舌鳎黏蛋白MUC5AC基因的克隆与结构预测

谭静, 胡秀彩, 吕爱军^通信作者, 孙敬锋, 刘小雪, 廉正义

天津农学院水产学院天津市水产生态及养殖重点实验室,天津 300384

收稿日期:2019-01-27 出版日期:2019-06-20 发布日期:2019-09-19
通讯作者: 吕爱军（1973-）,男,教授,博士,主要从事水产动物微生物与免疫学的研究。E-mail：lajand@126.com。
作者简介:谭静（1995-）,女,硕士在读,主要从事水产动物微生物与免疫学的研究。E-mail：1109849082@qq.com。
基金资助:
天津市自然科学基金重点项目（18JCYBJC29900,16JCZDJC33500）; 天津市水产生态及养殖重点实验室开放基金项目（TJAE201803）; 天津市水产产业技术体系创新团队（ITTFRS2017009）; 天津农学院综投平台项目（J01009030638）

Cloning and sequence analysis of mucin MUC5AC gene from the half-smooth tongue sole, Cynoglossus semilaevis

TAN Jing, HU Xiu-cai, LÜ Ai-jun, SUN Jing-feng, LIU Xiao-xue, LIAN Zheng-yi

Tianjin Key Lab of Aqua-Ecology and Aquaculture, College of Fisheries, Tianjin Agricultural University, Tianjin 300384, China

Received:2019-01-27 Online:2019-06-20 Published:2019-09-19

摘要/Abstract

摘要： 黏蛋白作为鱼类黏液的主要成分,在抵御病原感染过程中发挥重要作用。以半滑舌鳎（Cynoglossus semilaevis）黏蛋白为研究对象,采用RT-PCR方法对其皮肤黏蛋白MUC5AC（即CsMUC5AC）进行基因克隆及结构预测分析。结果获得CsMUC5AC基因片段的长度为2 771 bp,推测编码917个氨基酸,预测所含5个抗原表位分布在314-321、507-516、578-589、748-762和811-826 aa,3个结构域主要为C8（164-238,624-698 aa）、VWD（1-128,430-588 aa）和VWC（303-365,768-836 aa）。二级结构预测C8区主要由α-螺旋构成、VWD区以β折叠为主,三级结构蛋白亚基主要由VWD-C8-VWC组成。构建系统发育树分析显示,其与大菱鲆、攀鲈、罗非鱼黏蛋白MUC5AC亲缘关系较近,与分泌型黏蛋白家族自然聚为一支。以上结果为研究半滑舌鳎黏蛋白MUC5AC基因功能提供科学参考。

关键词: 半滑舌鳎, 黏蛋白, 基因克隆, 结构预测

Abstract: As the main component of fish mucus, mucin plays an important role in protecting from pathogen infection. The MUC5AC gene of Cynoglossus semilaevis, namely CsMUC5AC gene was cloned by RT-PCR method and protein structure was predicted by bioinformatics analysis. Results showed that the length of the CsMUC5AC gene fragment was 2 771 bp, encoding 917 amino acids（aa）, and 5 antigenic epitopes were predicted to be mainly located within 314-321, 507-516, 578-589, 748-762 and 811-826 aa sites. The 3 domains including C8（164-238, 624-698 aa）, VWD（1-128, 430-588 aa）and VWC（303-365, 768-836 aa）were observed. Analysis of secondary structure showed that the C8 motif was α-helix, the VWD domain was β-sheet, and the tertiary structure was two subunits profiles（VWD-C8-VWC）. The phylogenetic tree analysis showed that CsMUC5AC was closely related to that of Scophthalmus maximus, Anabas testudineus and Oreochromis niloticus, and naturally clustered into one group with the secretory mucins. These results provide a scientific reference for the further study of MUC5AC gene function in tongue sole.

Key words: Cynoglossus semilaevis, MUC5AC, gene cloning, structure prediction

中图分类号:

S917.4

谭静, 胡秀彩, 吕爱军, 孙敬锋, 刘小雪, 廉正义. 半滑舌鳎黏蛋白MUC5AC基因的克隆与结构预测[J]. 天津农学院学报, 2019, 26(2): 51-56.

TAN Jing, HU Xiu-cai, LÜ Ai-jun, SUN Jing-feng, LIU Xiao-xue, LIAN Zheng-yi. Cloning and sequence analysis of mucin MUC5AC gene from the half-smooth tongue sole, Cynoglossus semilaevis[J]. Journal of Tianjin Agricultural University, 2019, 26(2): 51-56.

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半滑舌鳎黏蛋白MUC5AC基因的克隆与结构预测

Cloning and sequence analysis of mucin MUC5AC gene from the half-smooth tongue sole, Cynoglossus semilaevis

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