CAJ | 학술논문

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 ）시일충인종류억제인자，기N단1-22위안기산가이휴대약물단백진입세포발휘약리작용，기위ARF。재시종단백EGFP화ARF간인입료이태（ linker 2）、유성칠태（ linker 7F）화강성칠태（linker 7S），연구기대융합단백EGFP-ARF천막활성적영향。실험결과현시인입칠태（linker 7）적융합단백，기형광강도고우linker 2적융합단백。격광공취초관찰표명，인입linker 7F적융합단백비인입linker 2적융합단백적천막효솔고5배좌우，비인입linker 7S적융합단백고3．8배좌우。결과표명련접태적형식대융합단백적결구화공능도유현저영향，위ARF작위약물운수재체적연구제공료의거。
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 .