Genetic Engineering Research Center, College of Bioengineering, Chongqing University, Chongqing 400030, People's Republic of China.
Biotechnol Lett. 2012 Mar;34(3):557-62. doi: 10.1007/s10529-011-0805-3. Epub 2011 Nov 23.
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.
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。