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| cbfa1、satb2双基因共表达载体的构建及鉴定 | |
| 引用本文： | Chen XX,Zhang J,Lv CX,Sun QF. cbfa1、satb2双基因共表达载体的构建及鉴定[J]. 上海口腔医学, 2011, 20(5): 474-478 |
| 作者姓名： | Chen XX Zhang J Lv CX Sun QF |
| 作者单位： | 1. 山东大学口腔医学院牙周科,山东,济南,250012 2. 山东省口腔生物医学重点实验室,山东,济南,250012 3. 山东大学口腔医学院牙周科山东省口腔生物医学重点实验室,山东,济南,250012 |
| 基金项目： | 山东省科技攻关项目(2009GG10002052)~~ |
| 摘 要： | 目的:构建携带cbfa1和satb2双基因的慢病毒真核表达载体。方法:采用PCR技术,从质粒中扩增基因cbfa1和satb2,经TA克隆、酶切筛选后,对阳性重组子进行商业测序鉴定。将测序正确的cbfa1和satb2分别插入pIRES上、下游的相应酶切位点中,构建pIRES-cbfa1-satb2。然后通过双酶切,得到cbfa1-Ires-satb2片段,将其插到含有neo/kana基因的慢病毒载体pLentinTrident1-CMV的相应酶切位点上,构建慢病毒真核表达载体pLentinTrident1-CMV-cbfa1-Ires-satb2。结果:通过PCR技术成功扩增了cbfa1和satb2基因,借助中间载体pIRES中的内部核糖体进入位点,将cbfa1和satb2同时连到了慢病毒载体pLentinTrident1-CMV上,经PCR、酶切及测序验证,质粒构建成功。结论:本实验成功构建了携带cbfa1和satb2双基因的慢病毒真核表达载体,为进一步研究2种基因的功能奠定了基础。 |
| 关 键 词： | Cbfa1 Satb2 慢病毒载体 |
Construction and identification of cbfa1 and satb2 co-expression vector |
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| Chen Xiao-xia,Zhang Jin,Lv Chun-xu,Sun Qin-feng. Construction and identification of cbfa1 and satb2 co-expression vector[J]. Shanghai journal of stomatology, 2011, 20(5): 474-478 | |
| Authors: | Chen Xiao-xia Zhang Jin Lv Chun-xu Sun Qin-feng |
| Affiliation: | CHEN Xiao-xia1,ZHANG Jin2,LV Chun-xu1,SUN Qin-feng1,2.(1.Department of Periodontics,College of Stomatology,Shandong University,2.Shandong Provincial Key Laboratory of Oral Biomedicine.Jinan 250012,Shandong Province,China) |
| Abstract: | PURPOSE: To construct a lentiviral eukaryotic expression vector containing two genes of cbfa1 and satb2.METHODS:The aim genes of cbfa1 and satb2 were amplified from plasmids by PCR.After TA cloning,the positive clones were identified by restrictive enzyme digestion and commercial DNA sequencing.Cbfa1 and satb2 with correct sequences were ligated upstream and downstream to pIRES,respectively,to construct pIRES-cbfa1-satb2.Then cbfa1-Ires-satb2 fragment was obtained by double digestion,and inserted into corre... |
| Keywords: | Cbfa1 Satb2 Lentivirus |
| 本文献已被 CNKI 万方数据 PubMed 等数据库收录！ | |