[沙蚕β-肌动蛋白基因的克隆及其内参作用]。

中药材 Pub Date : 2016-09-01
Yuan Li, Ai-li Zhang, Guo-dong Li, Zi-gang Qian
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引用次数: 0

摘要

目的:建立一种实时定量PCR检测沙梅烯囊膜蛋白β-肌动蛋白的方法,为qpcr检测沙梅烯囊膜蛋白基因提供参考基因。方法:根据β-actin基因保守区(genbank)设计特异性引物,采用PCR扩增β-actin基因。以β-actin为内参基因,采用q - PCR方法分析了沙参根、茎、叶中糖基转移酶基因(UGT)的表达。结果:沙草β-肌动蛋白基因扩增子长度为153 bp,与牛痘、水草和马齿苋β-肌动蛋白具有较高的同源性。以β-肌动蛋白为内参基因,发现UGT在沙生草不同组织中稳定表达。结论:β-肌动蛋白是研究三萜皂苷生物合成相关基因表达的可靠、合适的内参基因。
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[Cloning of β-actin Gene in Psammosilene tunicoides and Its Role as a Reference Gene].

Objective: To establish a real-time quantitative PCR method to detect Psammosilene tunicoides β-actin, and to provide a reference gene for the detection of Psammosilene tunicoides genes by q PCR.

Methods: Specific primers were designed based on the conserved region of the β-actin gene( Gen Bank) and were used to amplify β-actin by PCR. β-actin was also used as a reference gene in the q PCR analysis of glycosyltransferase gene( UGT) expression in the roots,stems,and leaves of Psammosilene tunicoides.

Results: The length of the β-actin gene amplicon from Psammosilene tunicoides was 153 bp and shared relatively high homology with β-actin found in Vaccaria segetalis, Myosoton aquaticum and Portulaca oleracea. Furthermore, UGT was revealed to be stably expressed in different Psammosilene tunicoides tissues when β-actin was employed as the reference gene.

Conclusion: β-actin is a reliable and suitable reference gene for studies on the expression of triterpenoid saponin biosynthesis-related genes in Psammosilene tunicoides.

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来源期刊
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11364
期刊介绍: "Chinese Materia Medica" magazine was founded in 1978. It is a national-level academic journal of Chinese medicine science and technology approved by the State Science and Technology Commission and the Press and Publication Administration for public distribution at home and abroad. It is sponsored by the Chinese Materia Medica Information Center of the State Food and Drug Administration and is publicly distributed at home and abroad. It mainly reports on new technologies, new methods and new achievements in the production, scientific research and business management of Chinese medicine, and serves to promote the production and scientific and technological development of Chinese medicine, improve the output and quality of Chinese medicine, improve business management, and prosper the Chinese medicine industry.
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