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| 人Delta-like4ext-26-217蛋白原核表达载体的构建及表达 | |
| 引用本文： | 黄斯勇.人Delta-like4ext-26-217蛋白原核表达载体的构建及表达[J].科学技术与工程,2011(22). |
| 作者姓名： | 黄斯勇 |
| 作者单位： | 1. 第四军医大学唐都医院血液科,西安,710038 2. 第四军医大学唐都医院血液科传染科,西安,710038 3. 第四军医大学基础部医学遗传学与发育生物学教研室,西安,710033 |
| 基金项目： | 国家自然科学基金面上项目(项目编号:30871090) |
| 摘 要： | 构建人Delta-like4ext-26-217原核表达载体并表达。采用PCR方法扩增hDll4ext-26-217多核苷酸序列，克隆入pMD18T载体。测序正确后，将其亚克隆入pET32a( )原核表达载体，获得pET32a( )-hDll4ext-26-217载体。以该载体转化E.coli菌株BL21，IPTG诱导其表达。采用SDS-PAGE、Western Blot方法鉴定目的蛋白的表达。结果表明，采用SDS-PAGE，Western Blot等方法均可检测到目的蛋白TRX/hDll4ext -26-217，分子量约42.2 KD，主要以包涵体形式大量存在。以上结果为Detla-like4功能的进一步研究奠定了基础。 |
| 关 键 词： | Detla-like4 Notch信号途径 原核表达 血管发生 |
| 收稿时间： | 4/27/2011 5:15:56 PM |
| 修稿时间： | 2011/4/28 0:00:00 |
Construction and Expression of human Delta-like4ext-26-217 fusion protein prokaryotic expression vector |
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| Hunag Si-Yong.Construction and Expression of human Delta-like4ext-26-217 fusion protein prokaryotic expression vector[J].Science Technology and Engineering,2011(22). | |
| Authors: | Hunag Si-Yong |
| Institution: | HUANG Si-yong1,WU Yan-lan2,LIU Li1,LI Guo-hui1,YIN Dan-dan1,HE Fei3,ZHANG Ping3,HAN hua3,LIANG Ying-min1*(Department of Hematology1,Department of Infectious Disease2,Tangdu Hospital,Fourth Military Medical University,Xi'an 710038,P.R.China,Department of Medical Genetics and Developmental Biology,Fourth Military Medical University3,Xi'an 710033,P.R.China) |
| Abstract: | To Construction and expression of human TRX/hDll4ext -26-217 fusion protein prokaryotic expression vector. The cDNA sequence of hDll4ext -26-217 was amplified by PCR, and cloned into the vector pMD18T. After sequencing, the correct fragment was cloned into prokaryotic expression plasmid pET32a( ). The target protein was expressed in E.coli BL21 induced by IPTG and the expression was detected by SDS-PAGE and Western blotting. The fusion protein TRX/hDll4ext -26-217 was detected by SDS-PAGE and Western blotting, and mainly aggregated into the inclusion body, which will be useful for further research on the function of Detla-like4. |
| Keywords: | Detla-like4 Notch signaling pathway prokaryotic expression angiogenesis |
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