| 摘要: |
| 一种从食用真菌Agrocybe aegerita中提取的凝集素AAL具有显著的抗肿瘤效果.该蛋白的基因序列已经通过RT-PCR得到.T7噬菌体展示库筛选发现该蛋白可以在体外和细胞核内的细胞周期调控蛋白mortality factor related gene on chromosome 15(MRG15)发生相互作用.本实验目的是证实AAL和MRG15在细胞核内的共定位,为AAL与MRG15的体内的互作提供证据.构建质粒,将aal与mrg15分别连接到pEGFP-C1和pDsRed-C1上,共转染Hela细胞.将pEGFP-C1和pD-sRed-C1空载体共转染Hela细胞作为对照.利用激光共聚焦显微镜研究AAL和MRG15在细胞内的共定位.AAL和MRG15均定位在细胞核内,荧光图片的重叠表明AAL和MRG15在核内共定位.而对照组中的EGFP和DS-RED在细胞内呈弥散分布.这些为AAL和MRG15在细胞核内的相互作用提供了证据. |
| 关键词: 凝集素 抗肿瘤 凋亡 共定位 |
| DOI： |
| 分类号:Q51 |
| 基金项目: |
|
| Co- localization of Antitumor Protein AAL and Target Protein MRG15 |
|
|
| Abstract: |
| An antitumor lectin from Agrocybe aegerita(AAL) has been purified and its cDNA sequence has been obtained through RT-PCR.T7 phage display library selection shows that AAL can interact with MRG15(mortality factor related gene on chromosome 15),an important cell cycle regulation factor in vitro.This study is toprovide evidence that AAL can interact with MRG15 in vivo through co-localization of these two proteins.pEGFP-aal and pDsRed-mrg15 plasmids were constructed and co-transfected into Hela cells for expression of both EGFP-AAL and DSRED-MRG15 fusion proteins,in order to determine the co-localization of AAL and MRG15 by confocal microscopy.Co-transfection of pEGFP-C1 and pDsRed-C1 empty vector plasmids were performed as control.Both AAL and MRG15 localized and aggregated in the same regions of the nucleus,and the merged image showed that both proteins co-localized,while in control both EGFP and DSRED proteins dispersed throughout the cell.This result provided evidence that AAL could interact with MRG15 in vivo. |
| Key words: lectin antitumor apoptosis co-localization |
