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来自秀丽隐杆线虫的一种经过修饰的n-3脂肪酸去饱和酶基因在转基因小鼠中产生了高比例的二十二碳六烯酸(DHA)和二十二碳五烯酸(DPA)。

A modified n-3 fatty acid desaturase gene from Caenorhabditis briggsae produced high proportion of DHA and DPA in transgenic mice.

作者信息

Zhu Guiming, Chen Hongxing, Wu Xiaojie, Zhou Yanrong, Lu Jianshen, Chen Hong, Deng Jixian

机构信息

Beijing Institute of Biotechnology, Beijing, China.

出版信息

Transgenic Res. 2008 Aug;17(4):717-25. doi: 10.1007/s11248-008-9171-x. Epub 2008 Mar 6.

Abstract

The functions of polyunsaturated fatty acids (PUFAs) have been widely investigated. In mammals, levels of n-3 PUFAs are relatively low compared to those of n-6 PUFAs. Either a lack of n-3 PUFAs or an excess of n-6 PUFAs could potentially cause health problems in humans. Hence, methods to increase the amount of n-3 PUFAs in diet have been intensely sought. In this study, we demonstrated that the n-3 fatty acid desaturase gene (sFat-1) synthesized from revised and optimized codons based on roundworm Caenorhabditis briggsae genomic gene for enhanced expression in mammals was successfully expressed in Chinese hamster ovary (CHO) cells and significantly elevated cellular n-3 PUFA contents. We generated sFat-1 transgenic mice by introducing mammal expression vector DNAs containing the sFat-1 gene into regular mice through the method of microinjection. Fatty acid compositions were then altered and the levels of docosahexaenoic acid (DHA, 22:6n-3) and docosapentaenoic acid (DPA, 22:5n-3) were greatly increased in these transgenic mice. Various types of tissues in the transgenic mice produced many types of n-3 PUFAs, such as alpha-linolenic acid (ALA; 18:3n-3), eicosapentaenoic acid (EPA, 20:5n-3), DPA, and DHA, for example, muscle tissues of the transgenic mice contained 12.2% DHA, 2.0% DPA, and 23.1% total n-3 PUFAs. These research results demonstrated that the synthesized sFat-1 gene with modified and optimized codons from C. briggsae possess functional activity and greater capability of producing n-3 PUFAs, especially DHA and DPA, in transgenic mice.

摘要

多不饱和脂肪酸(PUFAs)的功能已得到广泛研究。在哺乳动物中,与n-6多不饱和脂肪酸相比,n-3多不饱和脂肪酸的水平相对较低。n-3多不饱和脂肪酸的缺乏或n-6多不饱和脂肪酸的过量都可能对人类健康造成潜在问题。因此,人们一直在积极寻找增加饮食中n-3多不饱和脂肪酸含量的方法。在本研究中,我们证明,基于秀丽隐杆线虫基因组基因对密码子进行修订和优化后合成的n-3脂肪酸去饱和酶基因(sFat-1),用于在哺乳动物中增强表达,已在中国仓鼠卵巢(CHO)细胞中成功表达,并显著提高了细胞内n-3多不饱和脂肪酸的含量。我们通过显微注射法将含有sFat-1基因的哺乳动物表达载体DNA导入正常小鼠,从而培育出sFat-1转基因小鼠。这些转基因小鼠的脂肪酸组成发生了改变,二十二碳六烯酸(DHA,22:6n-3)和二十二碳五烯酸(DPA,22:5n-3)的水平大幅增加。转基因小鼠的各种组织都能产生多种类型的n-3多不饱和脂肪酸,如α-亚麻酸(ALA;18:3n-3)、二十碳五烯酸(EPA,20:5n-3)、DPA和DHA,例如,转基因小鼠的肌肉组织中含有12.2%的DHA、2.0%的DPA和23.1%的总n-3多不饱和脂肪酸。这些研究结果表明,从秀丽隐杆线虫中合成的经过修饰和优化密码子的sFat-1基因在转基因小鼠中具有功能活性,并且具有更强的产生n-3多不饱和脂肪酸的能力,尤其是DHA和DPA。

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