| 李斌元,何淑雅,王桂良,马云,孙春莉,肖卫纯,李洁,闵凌峰,彭丹妮,NanbertZhong.FXR2酵母双杂交饵载体的构建及酵母菌转化.[J].中南医学科学杂志.,2005,(3):299-302. |
| FXR2酵母双杂交饵载体的构建及酵母菌转化 |
| Construction and Transformation of Yeast Two-hybrid System "Bait" of FXR2 |
| 修订日期：2005-05-09 |
| DOI： |
| 中文关键词: 脆性X综合征 FXR2基因 酵母双杂交 诱饵质粒 EGY48（p8op—lacZ） |
| 英文关键词:fragile X syndrome,FXR2 gene,yeast two-hybrid system,Bait plasmid,EGY48(p8op-lacZ) |
| 基金项目:国家自然科学基金(30370795)，湖南省卫生厅基金(Y02-072). |
| 李斌元 何淑雅 王桂良 马云 孙春莉 肖卫纯 李洁 闵凌峰 彭丹妮 NanbertZhong |
| [1]南华大学生化与分子生物学研究室,湖南衡阳421001 [2]纽约州立基础研究所人类遗传研究室,美国纽约NY10314 |
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| 中文摘要: |
| 目的构建FXR2酵母双杂交系统的“饵”质粒，并转化酵母菌，为筛选FXR2P互作蛋白作基础。方法提取pMD18-T—FXR2，酶切后将FXR2基因连接到MATCHMAKER LexA Two—Hybrid System中的plexA质粒上，得到“诱饵”(Bait)-plexA—FXR2，导入大肠杆菌扩增，筛选阳性克隆并提取重组Bait质粒，再转入酵母菌EGY48(p8op—lacZ)。结果通过遗传学方法筛选得到构建成功的plexA—FXR2；通过营养缺陷筛选，证明Bait重组质粒转入酵母菌中。结论成功构建Bait质粒并使酵母菌转化。 |
| 英文摘要: |
| Objective Fragile X syndrome is a form of hereditary mental retardation, usually results from lack of expression of the FMR1 gene (fragile X mental retardation 1).There are two autosomal homologues of FMR1, called FXR1 and FXR2 (fragile X-related gene 1 and 2) .In this paper we aim to construct bait of yeast two-hybrid system,and then transform it into yeast in order to screen the interactive protein with FXR2P. Methods Distill pMD18-T vector.,then slice off FXR2 from pMD18-T -FXR2 and clone it into plexA of MATCHMAKER LexA Two-Hybrid System. After getting (Bait)- plexA- FXR2,amplify it through transforming into E.coli.screen for positive clones and distill recombined Bait plasmid.at the last transform into yeast. Results FXR2 was confirmed had been constructed into expressing vector plex a successfully by genetics means;It is proved that Bait has been transformed into yeast successifully through nutrition absence substrate. Conclusions Bait has been constructed and transformed into yeast. |
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