| 靶向人SSH1L的siRNA真核表达载体的构建及鉴定 |
| |
| 引用本文： | 邢志军,赵建武,王岩,高忠礼. 靶向人SSH1L的siRNA真核表达载体的构建及鉴定[J]. 中国实验诊断学, 2009, 13(1): 12-14 |
| |
| 作者姓名： | 邢志军 赵建武 王岩 高忠礼 |
| |
| 作者单位： | 1. 吉林大学中日联谊医院,吉林,长春,130033
2. 吉林大学病理生物学教育部重点实验室,吉林,长春,130021 |
| |
| 基金项目： | 吉林省科技厅白求恩专项基金，吉林省省科技厅国际合作项目，国家自然科学基金 |
| |
| 摘 要： | 目的构建靶向人SSH1L的siRNA真核表达载体（pSUPER-SSH1L）。方法将设计好的SSH1L的siRNA的寡聚脱氧核苷酸链与真核表达载体pSUPER连接，构建重组pSUPER-SSH1L真核表达载体。对该重组表达载体进行酶切鉴定及DNA测序；通过Westernblot检测SSH1L蛋白的表达。结果酶切鉴定、DNA测序以及Westernblot检测结果证实表达质粒构建成功，无碱基突变。结论成功构建了靶向人SSH1L基因表达的siRNA干扰质粒载体pStrPEn-SSH1L，为进一步运用RNA干扰技术进行SSH1L基因功能研究奠定了基础。
|
| 关 键 词： | SSH1L siRNA 真核表达载体 |
Construction and identification of eukaryotic expression vector of SSH1L-targeted siRNA |
| |
| Affiliation: | XING Zhi-jun, ZHAO Jian-wu,WANG Yan, et al. (1. China-Japan Union Hospital of Jilin University, Changchun 130033, China;2. The Key laboratory of Pathobiology of Chinese Ministry of Education, Jilin University, Changchun, Jilin 130021, China) |
| |
| Abstract: | Objective To construct a recombinant eukaryotic expression piasmid carrying human SSH1L-siRNA gene.Methons To connect the designed siRNA ＇ oligodeoxynucleofide chain of SSH1L with eukaryofic expression vector pSUPER, construct a recombinant eukaryotic expression plasmid pSUPER- SSH1L. Then to identify the recombinant expression plasmid by enzyme analysis and carry out DNA sequencing;and to check the expression of SSH1L protein through Western blot.Results The result of enzyme analysis,DNA sequencing and Western blot confirmed that the eukaryotic expression vector pSUPER-SSH1L was successfully con- structed,no basic radical mutation. Conclusion Eukaryotie expression vector pSUPER-SSH1L was successfully constructed, which was the basis for further studying SSH1L gene＇funcfion operating RNAi technique. |
| |
| Keywords: | SSH1L siRNA |
| 本文献已被 维普 万方数据 等数据库收录！ |
|
