文献类型：期刊文献

中文题名：一种人源性穿膜肽融合蛋白中连接肽的引入及其在HeLa细胞中的穿膜活性

英文题名：Introduction of Linking Peptides to a Human-derived Cell-penetrating Peptide and Its Transduction Activity in HeLa

作者：李堰忠[1];杨旭中[1];吕强[1];王富军[2];赵健[1]

机构：[1]华东理工大学生物反应器工程国家重点实验室,上海200237;[2]上海中医药大学,上海201203

年份：2013

卷号：35

期号：6

起止页码：1272

中文期刊名：江西农业大学学报

外文期刊名：Acta Agriculturae Universitatis Jiangxiensis

收录：CSTPCD;;北大核心:【北大核心2011】；CSCD:【CSCD_E2013_2014】；

基金：国家自然科学基金项目(81171449)

语种：中文

中文关键词：连接肽;ARF;融合蛋白;穿膜肽;重组表达

外文关键词：linker peptide;ARF ;fusion protein;cell-penetrating peptide;recombinant and expression

摘要：p14ARF(alternate reading frame)是一种人肿瘤抑制因子,其N端1-22位氨基酸可以携带药物蛋白进入细胞发挥药理作用,记为ARF。在示踪蛋白EGFP和ARF间引入了二肽(linker 2)、柔性七肽(linker 7F)和刚性七肽(linker 7S),研究其对融合蛋白EGFP-ARF穿膜活性的影响。实验结果显示引入七肽(linker 7)的融合蛋白,其荧光强度高于linker 2的融合蛋白。激光共聚焦观察表明,引入linker 7F的融合蛋白比引入linker 2的融合蛋白的穿膜效率高5倍左右,比引入linker 7S的融合蛋白高3.8倍左右。结果表明连接肽的形式对融合蛋白的结构和功能都有显著影响,为ARF作为药物运输载体的研究提供了依据。
p14ARF （alternate reading frame） is a human tumor inhibition factor .The first 22 amino acids, denoted by ARF ,can be absorbed by cells and then exert its function .In this study both of a 2 amino acids lin-ker peptide （ linker 2） ,a stable and flexible linking peptide of 7 amino acids （ linker 7S,linker 7F） were in-troduced to the site between Enhanced Green Fluorescence Peptide （EGFP）,a tracer peptide and ARF.The effect of three different linkers on recombinant protein EGFP-ARF was investigated .The results show that fluo-rescence intensity of proteins with 7 amino acids linker （ linker 7） is higher than that of linker 2,which means the length of linking peptide remarkably affects the function of EGFP .In virtue of laser confocal microscope ,the membrane-crossing activity is gained with the increase of linker length .Among this ,the uptake efficiency of re-combinant protein with linker 7F is 3.8 times higher than that of stable linking peptide and 5 times higher than that of linker 2.This study supplies a preferable construction method for using ARF as a valid carrier .