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A Novel Omega-3 Fatty Acid Desaturase Involved in Acclimation Processes of Polar Condition from Antarctic Ice Algae Chlamydomonas sp ICE-L

机译:涉及北极冰藻衣藻衣藻ICE-L极性条件驯化过程的新型Omega-3脂肪酸去饱和酶

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摘要

The ability of Antarctic ice algae, Chlamydomonas sp. ICE-L, to survive and proliferate at low temperature and high salinity implies that they have overcome key barriers inherent in Antarctic environments. A full-length complementary DNA (cDNA) sequence of omega-3 fatty acid desaturase, designated CiFAD3, was isolated via reverse transcription-polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends methods. The full-length of CiFAD3 cDNA contained an open reading frame of 1,302 bp with 5'-terminal untranslated region (UTR) of 36 bp and 3'-terminal UTR of 507 bp encoding a fatty acid desaturase protein of 434 amino acids. Sequence alignment and phylogenetic analysis showed that the gene was homologous to known chloroplastic omega-3 fatty acid desaturase. Meanwhile, CiFAD3 sequence showed typical features of membrane-bound desaturase such as three conserved histidine boxes along with four membrane spanning regions that were universally present among plant desaturases. Under different stress conditions, messenger RNA (mRNA) expression levels of CiFAD3 were measured by quantitative RT-PCR. The results showed that both temperature and salinity could motivate the upregulation of CiFAD3 expression. The mRNA accumulation of CiFAD3 increased 2.6-fold at 0A degrees C and 1.8-fold at 12A degrees C compared to the algae at 6A degrees C. Similarly, mRNA expression levels of CiFAD3 increased 3.8-fold after 62aEuro degrees NaCl treatment for 2 h. However, CiFAD3 mRNA expression levels were partially decreased after UV radiation. These data suggest that CiFAD3 is the enzyme responsible for the omega-3 fatty acid desaturation involved in ice algae Chlamydomonas sp. ICE-L acclimatizing to cold temperature and high salinity in Antarctic environment.
机译:南极冰藻衣藻的能力。 ICE-L在低温和高盐度下生存和增殖意味着它们已经克服了南极环境固有的主要障碍。通过逆转录-聚合酶链反应(RT-PCR)和cDNA末端快速扩增方法,分离出了ω-3脂肪酸去饱和酶的全长互补DNA(cDNA)序列,命名为CiFAD3。 CiFAD3 cDNA的全长包含一个1,302 bp的开放阅读框,其5'末端非翻译区(UTR)为36 bp,3'末端UTR为507 bp,编码434个氨基酸的脂肪酸去饱和酶蛋白。序列比对和系统发育分析表明,该基因与已知的叶绿体ω-3脂肪酸去饱和酶同源。同时,CiFAD3序列显示了膜结合的去饱和酶的典型特征,例如三个保守的组氨酸盒以及植物去饱和酶中普遍存在的四个跨膜区域。在不同的胁迫条件下,通过定量RT-PCR测量CiFAD3的信使RNA(mRNA)表达水平。结果表明,温度和盐度均可促进CiFAD3表达的上调。与6A摄氏度的藻类相比,CiFAD3的mRNA积累在0A摄氏度下增加了2.6倍,在12A摄氏度下增加了1.8倍。类似地,在62aEuro°NaCl处理2小时后,CiFAD3的mRNA表达水平增加了3.8倍。然而,CiFAD3 mRNA表达水平在紫外线照射后部分降低。这些数据表明,CiFAD3是负责冰藻衣藻(Chlamydomonas sp)中涉及的omega-3脂肪酸去饱和的酶。 ICE-L适应南极环境中的低温和高盐度。

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