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从茯苓中克隆和表征角鲨烯合酶基因及其被茉莉酸甲酯的上调。

Cloning and characterization of squalene synthase gene from Poria cocos and its up-regulation by methyl jasmonate.

机构信息

Department of Bioengineering, College of Food Science, South China Agricultural University, 482 Wu-Shan Road, Tian-He District, Guangzhou, 510642, Guangdong, China,

出版信息

World J Microbiol Biotechnol. 2014 Feb;30(2):613-20. doi: 10.1007/s11274-013-1477-z. Epub 2013 Sep 13.

Abstract

Squalene synthase (SQS) catalyzes the condensation of two molecules of farnesyl diphosphate to give presqualene diphosphate and the subsequent rearrangement to form squalene. The gene encoding squalene synthase was cloned from Poria cocos by degenerate PCR and inverse PCR. The open reading frame of the gene is 1,497 bp, which encodes 499 amino acid residues. A phylogenetic analysis revealed that P. cocos SQS belonged to the fungus group, and was more closely related to the SQS of Ganoderma lucidum than other fungi. The treatment of P. cocos with methyl jasmonate (MeJA) significantly enhanced the transcriptional level of P. cocos sqs gene and the content of squalene in P. cocos. The transcriptional level of sqs gene was approximately fourfold higher than the control sample and the squalene content reached 128.62 μg/g, when the concentration of MeJA was 300 μM after 72 h induction.

摘要

鲨烯合酶(SQS)催化两分子法呢基二磷酸缩合生成预鲨烯二磷酸,并随后重排形成鲨烯。通过简并 PCR 和反向 PCR,从茯苓中克隆出编码鲨烯合酶的基因。该基因的开放阅读框为 1497bp,编码 499 个氨基酸残基。系统发育分析表明,P. cocos SQS 属于真菌群,与灵芝的 SQS 比其他真菌更为密切相关。用茉莉酸甲酯(MeJA)处理茯苓可显著提高 P. cocos sqs 基因的转录水平和茯苓中鲨烯的含量。当 MeJA 浓度为 300μM 并诱导 72 小时后,sqs 基因的转录水平约为对照样品的 4 倍,鲨烯含量达到 128.62μg/g。

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