National Glycoengineering Research Center and College of Life Science, Shandong University, Jinan, 250100, People's Republic of China.
Mar Biotechnol (NY). 2011 Jun;13(3):393-401. doi: 10.1007/s10126-010-9309-8. Epub 2010 Jul 29.
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 0°C and 1.8-fold at 12°C compared to the algae at 6°C. Similarly, mRNA expression levels of CiFAD3 increased 3.8-fold after 62‰ 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.
南极冰藻 Chlamydomonas sp. ICE-L 能够在低温和高盐度下生存和繁殖,这意味着它们已经克服了南极环境中固有的关键障碍。通过逆转录-聚合酶链反应(RT-PCR)和快速扩增 cDNA 末端方法,分离出一种全长 ω-3 脂肪酸去饱和酶的 cDNA 序列,命名为 CiFAD3。CiFAD3 cDNA 的全长包含一个 1302bp 的开放阅读框,5'非翻译区(UTR)为 36bp,3'UTR 为 507bp,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,该基因与已知的质体 ω-3 脂肪酸去饱和酶同源。同时,CiFAD3 序列表现出典型的膜结合去饱和酶特征,如三个保守的组氨酸盒以及普遍存在于植物去饱和酶中的四个跨膜区。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,温度和盐度都能促使 CiFAD3 的表达上调。与在 6°C 下的藻类相比,CiFAD3 在 0°C 和 12°C 下的 mRNA 积累分别增加了 2.6 倍和 1.8 倍。同样,在 62‰NaCl 处理 2 小时后,CiFAD3 的 mRNA 表达水平增加了 3.8 倍。然而,CiFAD3 的 mRNA 表达水平在紫外线辐射后部分下降。这些数据表明,CiFAD3 是负责参与南极冰藻 Chlamydomonas sp. ICE-L 适应低温和高盐度的 ω-3 脂肪酸去饱和的酶。