Jin Xiaoli, Feng Dongru, Wang Hongbin, Wang Jinfa
The State Key Laboratory of Biocontrol and The Key Laboratory of Gene Engineering of Ministry of Education, School of Life Sciences, Sun Yat-Sen University, Guangzhou, 510275, China.
Biotechnol Lett. 2007 Sep;29(9):1431-7. doi: 10.1007/s10529-007-9403-9. Epub 2007 May 26.
A new full-length beta-1,3-glucanase cDNA, MpGlu, was isolated from a plantain (Musa paradisica) by the rapid amplification of cDNA ends (RACE) technique. Recombinant GST-MpGlu protein, expressed in E. coli, hydrolyzed (1-->3),(1-->6)-beta-glucan of Laminaria digitata and inhibited the growth of Fusarium oxysporum fsp. cubense (race 4) suggesting that it is a beta-1,3-glucanase. Southern blot analysis indicated that there is one copy of MpGlu in the plantain genome. MpGlu gene expression was detected in plantain leaves, peel, and pulp by RT-PCR. Northern blot analysis revealed that the expression of MpGlu was up-regulated by Fusarium infection. Subcellular localization analysis indicated that 28 residues at the N-terminal end are necessary for extracellular secretion, while 32 residues at the C-terminal end are necessary to target the protein into vacuoles.
通过cDNA末端快速扩增(RACE)技术从大蕉(Musa paradisica)中分离出一个新的全长β-1,3-葡聚糖酶cDNA,即MpGlu。在大肠杆菌中表达的重组GST-MpGlu蛋白可水解掌状海带的(1→3),(1→6)-β-葡聚糖,并抑制尖孢镰刀菌古巴专化型(4号生理小种)的生长,这表明它是一种β-1,3-葡聚糖酶。Southern杂交分析表明大蕉基因组中存在一个MpGlu拷贝。通过RT-PCR在大蕉的叶片、果皮和果肉中检测到MpGlu基因表达。Northern杂交分析显示,镰刀菌感染可上调MpGlu的表达。亚细胞定位分析表明,N末端的28个残基是细胞外分泌所必需的,而C末端的32个残基是将蛋白质靶向液泡所必需的。