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强启动子 PMagpd 为昆虫病原真菌绿僵菌中的高基因表达提供了工具。

A strong promoter, PMagpd, provides a tool for high gene expression in entomopathogenic fungus, Metarhizium acridum.

机构信息

Genetic Engineering Research Center, College of Bioengineering, Chongqing University, Chongqing 400030, People's Republic of China.

出版信息

Biotechnol Lett. 2012 Mar;34(3):557-62. doi: 10.1007/s10529-011-0805-3. Epub 2011 Nov 23.

Abstract

A glyceraldehyde-3-phosphate dehydrogenase gene (gpd) promoter (PMagpd) was obtained from Metarhizium acridum and its active region analyzed by 5'-deletion strategy using β-glucuronidase (GUS) as a reporter. Sequence analysis revealed that typical regulatory elements of PMagpd were included in the 1.7 kb region upstream of the start codon of the Magpd gene. Deletion of the region from -1,691 bp to -1,463 bp, where the gpd box is harbored, did not significantly affect the PMagpd activity. Deletions of the regions upstream of -946 bp and upstream of -684 bp caused a major decrease of GUS activity. Compared with PgpdA (2.2 kb) in Aspergillus nidulans, PMagpd (1.4 kb) had a shorter sequence and significantly higher activity in M. acridum. This study provides an applicable promoter for over-expression of target genes in M. acridum.

摘要

从蝗绿僵菌中获得了 1,3-二磷酸甘油醛脱氢酶基因(gpd)启动子(PMagpd),并通过 5'缺失策略用β-葡萄糖醛酸酶(GUS)作为报告基因对其活性区进行了分析。序列分析表明,在 Magpd 基因起始密码子上游 1.7kb 区域包含了 PMagpd 的典型调控元件。从-1691bp 到-1463bp 缺失 gpd 盒所在区域对 PMagpd 活性没有显著影响。-946bp 上游和-684bp 上游缺失区域的缺失导致 GUS 活性的显著下降。与棘孢木霉(Aspergillus nidulans)中的 PgpdA(2.2kb)相比,PMagpd(1.4kb)具有更短的序列,在蝗绿僵菌中具有更高的活性。本研究为在蝗绿僵菌中过表达目标基因提供了一种可行的启动子。

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