| 摘要: |
| DHA和EPA对心脑血管疾病以及癌症等疾病的防治有着重要的作用,其中ω-3和ω-6脂肪酸脱氢酶是合成多不饱和脂肪酸DHA和EPA过程中的两个关键的去不饱和酶,由于缺乏这两种酶的基因,高等动物无法自发合成多不饱和脂肪酸,为了得到可以生产多不饱和脂肪酸的转基因动物,本研究计划将大豆的这两个去饱和脂肪酸作为目的基因,但由于不同物种之间的密码子使用具有偏爱性,会严重影响到目的基因在宿主体内的表达水平,为了解决这一问题,提高目的基因在宿主体内的表达水平,同时采用Jcat在线软件和手动替换密码子两种方法对大豆的ω-3和ω-6脂肪酸脱氢酶基因进行了优化,然后选出最优的优化方法,为后期进一步的实验提供目的基因。最后本文预测了两种优化结果的RNA二级结构和CAI、CBI、Nc值和GC含量,比较得出手动替换的结果优于在线优化的结果。 |
| 关键词: 多不饱和脂肪酸 脂肪酸脱氢酶 密码子偏爱性 基因优化 转基因 |
| DOI：10.3969/j.issn.1672-5565.201707003 |
| 分类号:Q522+.6 |
| 文献标识码:A |
| 基金项目:国家自然科学基金(61561036);内蒙古自治区杰出青年培育基金(2017JQ04). |
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| The codon optimization of ω-3 and ω-6 fatty desaturase genes in Glycine max |
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JIA Xueqi, ZUO Yongchun
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(The Research Center for Laboratory Animal Science, College of Life Sciences, Inner Mongolia University, Hohhot 010070, China)
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| Abstract: |
| Docosahexaenoic acid (DHA) and Eicosapentaenoic acid (EPA) are of significance to prevent the cardiovascular diseases and cancers. ω-3 fatty desaturases and ω-6 fatty desaturases are two key enzymes in the biosynthetic pathway of polyunsaturated fatty acid. Because of the lack of the genes of these two enzymes, the mammals cannot synthesize PUFAs spontaneously. To gain the transgenic animals which can produce these PUFAs, this study plans to transfer the two genes into mammals’ bodies. However, the codon usage biases are different between different species, which can significantly affect the expression level of the target genes in hosts. To solve this problem and improve the expression level, two optimization methods, Jcat online optimization software and manual codon replacement are applied at the same time to optimize ω-3 fatty desaturase gene and ω-6 fatty desaturase gene of Glycine max and the best method will be used in the further experiments. At last, this study has predicted the RNA second structures and CAI, CBI, Nc and GC% of the two optimization results and the analysis shows that the manual optimization is better to improve the expression level. |
| Key words: Polyunsaturated fatty acids Fatty desaturases Codon bias Gene optimization Transgenesis |
