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引用本文:王坤,王沛,张浩,张树峰,王大志.米氏凯伦藻细胞表面膜蛋白质组及其对温度变化的响应研究.海洋与湖沼,2019,50(3):652-663.
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米氏凯伦藻细胞表面膜蛋白质组及其对温度变化的响应研究
王坤1, 王沛1, 张浩1, 张树峰1, 王大志1,2
1.厦门大学近海海洋环境科学国家重点实验室 厦门 361102;2.中国科学院海洋生态与环境科学重点实验室 青岛 266071
摘要:
米氏凯伦藻(Karenia mikimotoi)近年在我国福建、浙江和广东沿海经常形成赤潮,其赤潮不仅影响到海洋生态系统的稳定,也严重威胁到水产养殖以及人类生命健康安全。本论文以米氏凯伦藻为研究对象,建立了米氏凯伦藻细胞表面膜蛋白质荧光标记技术和细胞膜蛋白质提取方法,运用荧光差异凝胶电泳技术(2-D DIGE)对膜蛋白质进行了分析,并研究了米氏凯伦藻的膜蛋白质组及其对环境温度变动的响应。实验共鉴定到44个细胞表面膜蛋白,其中有效注释27个,主要为转运蛋白、HSP70蛋白家族和捕光蛋白等。米氏凯伦藻在20℃条件下的细胞生长和光合作用要明显好于16℃和12℃,但16℃和12℃条件下的差别不大,表明低温限制了米氏凯伦藻的生长。当米氏凯伦藻从12℃快速转移至16℃和20℃时,藻细胞密度和光合作用效率短时间迅速降低,但细胞很快即适应温度变化。细胞膜上的转运蛋白和光合作用蛋白在其适应温度变化中起着重要作用。
关键词:  米氏凯伦藻  温度  膜蛋白质  蛋白质组  差异荧光凝胶电泳技术
DOI:10.11693/hyhz20180700189
分类号:Q946;Q948.885.3;X55
基金项目:国家重点研发计划资助项目,2017YFC1404300号
THE CELL SURFACE PROTEOME OF KARENIA MIKIMOTOI AND ITS RESPONSE TO TEMPERATURE CHANGE
WANG Kun1, WANG Pei1, ZHANG Hao1, ZHANG Shu-Feng1, WANG Da-Zhi1,2
1.State Key Laboratory of Marine Environmental Science, Xiamen University, Xiamen 361102, China;2.CAS Key Laboratory of Marine Ecology and Environmental Sciences, Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071, China
Abstract:
Karenia mikimotoi is a toxic harmful algal bloom species distributed in the temperate and tropical coastal waters, and it can produce hemolytic toxin and fish toxin. In recent years, it often causes blooms in the coastal areas of Fujian, Zhejiang, and Guangdong provinces of China, resulting in the destabilization of the marine ecosystem and serious harms to local aquaculture and human health or even life. In this paper, fluorescence labeling and sequential extraction of cell membrane proteins from K. mikimotoi were established. Cell surface membrane proteins were analyzed using fluorescence difference gel electrophoresis (2-D DIGE), and the response of membrane proteome of K. mikimotoi to a changing environmental temperature was studied. In total, 44 cell surface membrane proteins were identified, of which 27 were annotated effectively, including mainly transporters, HSP70 protein family, and light harvesting protein. Cell growth and photosynthesis of K. mikimotoi grown in 20℃ were better than those in 16℃ and 12℃ but no difference was found between 16℃ and 12℃, indicating that a low temperature could limit the cell growth of K. mikimotoi. When ambient temperature of K. mikimotoi cells boosted rapidly from 12℃ to 16℃ or 20℃, cell density and photosynthesis efficiency decreased quickly but the cells could rapidly adapt to the temperature change. Therefore, we believe that the transport and photosynthetic proteins of a cell membrane play an important role in the cells' adaptation to a quick ambient temperature change.
Key words:  Karenia mikimotoi  temperature  membrane proteins  proteome  difference fluorescence gel electrophoresis
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