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[用于灵芝中(1,3)-β-葡聚糖合酶基因扩增及该酶部分特性研究的引物]

[Primers for (1,3)-β-glucan synthase gene amplification and partial characterization of the enzyme in Ganoderma lucidum].

作者信息

Guerrero-Torres Jessica Valeria, Mata Gerardo, Martínez-Carrera Daniel, Garibay-Orijel Claudio, Garibay-Orijel Roberto

机构信息

Departamento de Bioingeniería, Unidad Profesional Interdisciplinaria de Biotecnología, Instituto Politécnico Nacional, México, D. F., México.

出版信息

Rev Iberoam Micol. 2013 Oct-Dec;30(4):267-70. doi: 10.1016/j.riam.2012.12.006. Epub 2013 Feb 9.

Abstract

BACKGROUND

β-(1,3)(1,6)-D-glucan is fungal cell wall component that has demonstrated immunomodulatory and anti-cancer effects. The (1,3)-β-glucan synthase is one of the main enzymes involved in its biosynthesis.

AIMS

To design primers to partially amplify and characterize the (1,3)-β-glucan synthase gene and to determine them in Ganoderma lucidum (G. Lucidum) strain CP-132.

METHODS

The primers were designed on the basis of homologous genes in other fungi. Then, using the PCR technique, primers were tested using DNA extracted from the G. lucidum strain CP-382. Amplified sequences were compared with those from the GenBank.

RESULTS

Three primer pairs were designed; all of them produced amplicons of the expected size. The sequences obtained with primer pairs BGS2113UmF and BGS3097UmR, and BGS547UmF and BGS2113UmR matched with 2 sections of the (1,3)-β-glucan synthase gene. The deduced amino acid sequences showed high similarity with homologous genes from other fungi, particularly with those of the Agaricomycetes class.

CONCLUSIONS

The primer design to partially amplify the (1,3)-β-glucan synthase gene of G. lucidum using sequences from homologous genes was successful. These primers will allow to characterize this important enzyme in a wide group of fungi.

摘要

背景

β-(1,3)(1,6)-D-葡聚糖是真菌细胞壁的组成成分,已显示出免疫调节和抗癌作用。(1,3)-β-葡聚糖合酶是参与其生物合成的主要酶之一。

目的

设计引物以部分扩增和鉴定(1,3)-β-葡聚糖合酶基因,并在灵芝菌株CP-132中对其进行测定。

方法

根据其他真菌中的同源基因设计引物。然后,使用PCR技术,用从灵芝菌株CP-382中提取的DNA对引物进行测试。将扩增序列与来自GenBank的序列进行比较。

结果

设计了三对引物;所有引物均产生了预期大小的扩增子。引物对BGS2113UmF和BGS3097UmR以及BGS547UmF和BGS2113UmR获得的序列与(1,3)-β-葡聚糖合酶基因的2个区段匹配。推导的氨基酸序列与其他真菌的同源基因具有高度相似性,特别是与伞菌纲的同源基因。

结论

利用同源基因序列设计引物以部分扩增灵芝(1,3)-β-葡聚糖合酶基因的方法是成功的。这些引物将有助于在多种真菌中鉴定这种重要的酶。

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