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南极绿藻莱茵衣藻ICE-L中钙调蛋白的克隆及胁迫诱导表达分析

Cloning and Stress-Induced Expression Analysis of Calmodulin in the Antarctic Alga Chlamydomonas sp. ICE-L.

作者信息

He Ying-Ying, Wang Yi-Bin, Zheng Zhou, Liu Fang-Ming, An Mei-Ling, He Xiao-Dong, Qu Chang-Feng, Li Lu-Lu, Miao Jin-Lai

机构信息

The First Institute of Oceanography, State Oceanic Administration, No. 6 of Xianxialing Road, Qingdao, 266061, China.

Key Laboratory of Marine Bioactive Substances, SOA, Qingdao, 266061, China.

出版信息

Curr Microbiol. 2017 Aug;74(8):921-929. doi: 10.1007/s00284-017-1263-5. Epub 2017 May 17.

Abstract

Calmodulin (CaM) is a Ca-binding protein that plays a role in several Ca signaling pathways, which dynamically regulates the activities of hundreds of proteins. The ice alga Chlamydomonas sp. ICE-L, which has the ability to adapt to extreme polar conditions, is a crucial primary producer in Antarctic ecosystem. This study hypothesized that Cam helps the ICE-L to adapt to the fluctuating conditions in the polar environment. It first verified the overall length of Cam, through RT-PCR and RACE-PCR, based on partial Cam transcriptome library of ICE-L. Then, the nucleotide and predicted amino acid sequences were, respectively, analyzed by various bioinformatics approaches to gain more insights into the computed physicochemical properties of the CaM. Potential involvements of Cam in responding to certain stimuli (i.e., UVB radiation, high salinity, and temperature) were investigated by differential expression, measuring its transcription levels by means of quantitative RT-PCR. Results showed that CaM was indeed inducible and regulated by high UVB radiation, high salinity, and nonoptimal temperature conditions. Different conditions had different expression tendencies, which provided an important basis for investigating the adaptation mechanism of Cam in ICE-L.

摘要

钙调蛋白(CaM)是一种钙结合蛋白,在多种钙信号通路中发挥作用,可动态调节数百种蛋白质的活性。冰藻莱茵衣藻ICE-L能够适应极端极地条件,是南极生态系统中的关键初级生产者。本研究假设CaM有助于ICE-L适应极地环境中的波动条件。基于ICE-L的部分CaM转录组文库,首先通过RT-PCR和RACE-PCR验证了CaM的全长。然后,分别采用各种生物信息学方法分析核苷酸和预测的氨基酸序列,以更深入了解CaM的计算物理化学性质。通过差异表达,利用定量RT-PCR测量其转录水平,研究了CaM在响应某些刺激(即UVB辐射、高盐度和温度)中的潜在作用。结果表明,CaM确实可被高UVB辐射、高盐度和非最适温度条件诱导和调节。不同条件下有不同的表达趋势,这为研究CaM在ICE-L中的适应机制提供了重要依据。

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