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红螯螯虾(Cherax quadricarinatus)Δ9 脂肪酸去饱和酶样基因的克隆与特性及其在冷胁迫下的表达分析。

Cloning and characterisation of a Δ9 fatty acyl desaturase-like gene from the red claw crayfish (Cherax quadricarinatus) and its expression analysis under cold stress.

机构信息

Institute for Agri-food Standards and Testing Technology, Shanghai Academy of Agricultural Science, Shanghai, 201106, China.

Shanghai Runzhuang Agricultural Science and Technology Co., Ltd, China.

出版信息

J Therm Biol. 2021 Dec;102:103122. doi: 10.1016/j.jtherbio.2021.103122. Epub 2021 Nov 2.

Abstract

Desaturase is one of the key enzymes in the unsaturated fatty acid synthesis pathway. Δ9 desaturase catalyzes the synthesis of oleic acid from stearic acid by introducing double bonds in the 9th and 10th carbon chains, thereby increasing the content of MUFAs in the body. In order to explore the main function of the Δ9 desaturase gene under low temperature stress, RACE-PCR technology was used in this study to clone the full-length sequence of the CqFAD9-like from the hepatopancreas of red claw crayfish, Cherax quadricarinatus. The full length of the sequence is 1236 bp, and the open reading frame is 1041 bp, encoding 346 amino acid residues. The 5 'UTR is 116 bp, the 3' UTR is 79 bp, and the 3 'UTR contains a PloyA tail. The predicted theoretical isoelectric point and molecular weight are 8.68 and 40.28 kDa, respectively. Homology analysis showed that the sequence had the highest similarity with FAD9 from crustaceans. The results of real-time PCR showed that the expression level of this gene was highest in the hepatopancreas, which was significantly higher than other tissues, followed by the ovaries, brain ganglion and stomach. At the same time, the expression of the CqFAD9-like in hepatopancreas of crayfish cultured at 25, 20, 15 and 9 °C for four weeks was detected. The results showed that expression of the FAD9 gene increased gradually with decreasing temperature, indicating that metabolic desaturation might play a regulatory role during cold stress.

摘要

去饱和酶是不饱和脂肪酸合成途径中的关键酶之一。Δ9 去饱和酶通过在第 9 位和第 10 位碳链中引入双键,催化硬脂酸合成油酸,从而增加体内 MUFA 的含量。为了探索 Δ9 去饱和酶基因在低温胁迫下的主要功能,本研究采用 RACE-PCR 技术克隆了罗氏沼虾(Cherax quadricarinatus)肝胰腺中 CqFAD9-like 的全长序列。序列全长为 1236bp,开放阅读框为 1041bp,编码 346 个氨基酸残基。5'UTR 为 116bp,3'UTR 为 79bp,3'UTR 含有 PolyA 尾巴。预测的理论等电点和分子量分别为 8.68 和 40.28kDa。同源分析表明,该序列与甲壳动物的 FAD9 具有最高的相似性。实时 PCR 结果显示,该基因在肝胰腺中的表达水平最高,明显高于其他组织,其次是卵巢、脑和胃。同时,检测了在 25、20、15 和 9°C 下培养四周的罗氏沼虾肝胰腺中 CqFAD9-like 的表达。结果表明,FAD9 基因的表达随着温度的降低而逐渐增加,表明代谢去饱和可能在冷应激过程中发挥调节作用。

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