| 过量表达OsUgp2基因提高紫芝多糖含量 |
| Increasing polysaccharide yield by over-expression of OsUgp2 in Ganoderma sinense |
| |
| 中文关键词:尿苷二磷酸葡萄糖焦磷酸化酶，过量表达，灵芝，多糖 |
| 英文关键词:UDP-glucose pyrophosphorylase, over-expression, Ganoderma sinense, polysacchride |
| 基金项目:国家自然科学基金“水稻花药特异表达UDPG焦磷酸化酶基因的功能分析”（No. 30370800） |
|
| 摘要点击次数: 1936 |
| 全文下载次数: 1174 |
| 中文摘要: |
| 尿苷二磷酸葡萄糖焦磷酸化酶（UDP-glucose pyrophosphorylase，UGPase）是多糖生物合成过程中重要的酶，水稻基因组中存在两个UGPase同源基因分别命名为OsUgp1和OsUgp2。构建了由构巢曲霉3-磷酸甘油醛脱氢酶基因启动子驱动OsUgp2表达的真菌过量表达载体，并通过农杆菌介导法将OsUgp2基因转入紫芝中，获得了潮霉素抗性的转化菌株。PCR和Southern杂交结果显示OsUgp2基因成功整合到受体紫芝基因组中。半定量RT-PCR检测结果显示外源基因OsUgp2在紫芝转 |
| 英文摘要: |
| UDP-glucose pyrophosphorylase (UGPase) plays an important role in the process of polysaccharide synthesis. Two UGPase homologous genes existing in rice genome were designated as OsUgp1 and OsUgp2. In this study, an over-expression vector, in which OsUgp2 gene was driven by a promoter of glyceraldehyde-3-phosphate dehydrogenase from Aspergillus nidulans, was constructed and introduced into Ganoderma sinense via Agribacterium tumefaciens mediated method. A number of hygromycin resistant transformants was obtained. PCR and Southern blot analyses showed that the exogenous OsUgp2 was integrated into the genome of Ganoderma sinense. The result of semi-quantitative RT-PCR indicated that OsUgp2 expressed and the expression level was different among the three independent transformants. The determined UGPase activity of transformants was found increased and its extent was correlated well with the mRNA level. The constant weight and phenol-sulfuric acid assays were utilized to determine the mycelial biomass and polysaccharide content respectively. The results showed that all of biomass, intracellular and extracellular polysaccharides were evidently increased. The positive correlation between UGPase activity and polysaccharide content was revealed by analysis using SPSS13.0 software. The correlation coefficient between UGPase activity and intracellular polysaccharide was the highest at 0.980. It is suggested that the overexpressed OsUgp2 is probably more concerned with intracellular polysaccharide synthesis. |
| 张帆,钟威,穆虹,李刚. 过量表达OsUgp2基因提高紫芝多糖含量[J]. 菌物学报, 2011, 30(3): 442-452 |
| 查看全文 查看/发表评论 下载PDF阅读器 |
|