食品科学

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低温诱导球等鞭金藻3011脂肪酸去饱和酶基因片段的筛选及mRNA表达分析

王 婷,赵 培,李 楠,王雪青*   

  1. 天津商业大学生物技术与食品科学学院,天津市食品生物技术重点实验室,天津 300134
  • 出版日期:2016-03-15 发布日期:2016-03-17

mRNA Expression Analysis and Screening of Fatty Acid Desaturase Gene in Isochrysis galbana 3011 under Low-Temperature Induction

WANG Ting, ZHAO Pei, LI Nan, WANG Xueqing*   

  1. Tianjin Key Laboratory of Food Biotechnology, College of Biotechnology and Food Science,Tianjin University of Commerce, Tianjin 300134, China
  • Online:2016-03-15 Published:2016-03-17

摘要:

本实验通过设计特异引物序列,对经低温诱导处理的球等鞭金藻3011的脂肪酸去饱和酶基因片段进行筛选,以实时荧光定量聚合酶链式反应检测酶的表达量,探索低温对该多不饱和脂肪酸脱饱和酶基因表达量的影响。结果表明:用prime5设计包括内参基因在内的14 条引物,其中7 条引物具有特异性扩增,根据扩增效果对其扩增体系进行优化。结果表明:Des4、Des5、Des6、Des8、Des9和Des12 mRNA相对最大表达量分别为7.71、11.33、42.58、22.02、73.91和16.01。随温度的降低均呈现先增加后降低的趋势。根据SPSS 19.0统计分析,6 个基因在其mRNA相对表达量最大的诱导时间时,18 ℃的mRNA相对表达量与其在12、15、21 ℃ 3 个温度条件下的相对表达量相比具有极显著差异(P<0.01)。18 ℃条件下诱导24 h可以有效增加Des9基因在球等鞭金藻3011中的表达。此研究结果为人为调控微藻的多不饱和脂肪酸含量提供了一条可行的方法。

关键词: 低温, 脂肪酸去饱和酶, 球等鞭金藻3011, 实时荧光定量聚合酶链式反应, 基因表达

Abstract:

In the present study, the specific primers were designed for the screening of the fatty acid desaturase gene
in Isochrysis galbana 3011 under low-temperature induction and for the determination of fatty acid desaturase by real
time-qPCR. Results showed that of 14 primers designed using prime5, 7 provided specific amplification. Based on
amplification efficiency, the largest relative expression amounts of Des4, Des5, Des6, Des8, Des9 and Des12 were 7.71,
11.33, 42.58, 22.02, 73.91 and 16.01, respectively, all of which dropped after an initial rise with decreasing temperature.
Statistical analysis performed using SPSS19.0 software indicated that there was highly significant difference between the
relative expression amounts of 6 genes at 18 ℃ and those obtained at three other temperatures 12, 15, 21 ℃ (P < 0.01). The
induction at 18 ℃ for 24 h could effectively increase the expression of Des9 in Isochrysis galbana 3011. This study has
demonstrated a feasible method to manipulate the content of polyunsaturated fatty acids in microalgae.

Key words: low temperature, fatty acids desaturase, Isochrysis galbana 3011, real time fluorescence quantitative polymerase chain reaction(real time-qPCR), gene expression

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