[发明专利]一种人野生型P53融合蛋白

摘要

本发明涉及一种P53融合蛋白。本发明将人wtP53基因cDNA片段和蛋白传导结构域(PTD)融合，插入到大肠杆菌原核表达载体PGEX－3X中，得到重组表达质粒PGEX－3X－P53，利用IPTG诱导产生wtP53融合蛋白；分子量约为53kbp，经DNA序列测定，它由1218个核苷酸组成；经MTT法及TUMEL法体外实验，表明wtP53融合蛋白可明显抑制肺癌细胞H1299和肝癌细胞HePG2的增殖，促进其调亡，本发明为应用wtP53融合蛋白治疗肿瘤提供了可靠的途径。

主权项

1、一种人野生型P53融合蛋白，其特征在于该融合蛋白的cDNA的核苷酸序列和相应的编码氨基酸序列如下：ggg atc cgg cgt cgt cag cag cga cgt aaa aag agg atg gag gag ccg 48Gly Ile Arg Arg Arg Gln Gln Arg Arg Lys Lys Arg Met Glu Glu Pro1 5 10 15cag tca gat cct agc gtc gag ccc cct ctg agt cag gaa aca ttt tca 96Gln Ser Asp Pro Ser Val Glu Pro Pro Leu Ser Gln Glu Thr Phe Ser 20 25 30gac cta tgg aaa cta ctt cct gaa aac aac gtt ctg tcc ccc ttg ccg 144Asp Leu Trp Lys Leu Leu Pro Glu Asn Asn Val Leu Ser Pro Leu Pro 35 40 45tcc caa gca atg gat gat ttg atg ctg tcc ccg gac gat att gaa caa 192Ser Gln Ala Met Asp Asp Leu Met Leu Ser Pro Asp Asp Ile Glu Gln 50 55 60tgg ttc act gaa gac cca ggt cca gat gaa gct ccc aga atg cca gag 240Trp Phe Thr Glu Asp Pro Gly Pro Asp Glu Ala Pro Arg Met Pro Glu65 70 75 80gct gct ccc ccc gtg gcc cct gca cca gca gct cct aca ccg gcg gcc 288Ala Ala Pro Pro Val Ala Pro Ala Pro Ala Ala Pro Thr Pro Ala Ala 85 90 95cct gca cca gcc ccc tcc tgg ccc ctg tca tct tct gtc cct tcc cag 336Pro Ala Pro Ala Pro Ser Trp Pro Leu Ser Ser Ser Val Pro Ser Gln 100 105 110aaa acc tac cag ggc agc tac ggt ttc cgt ctg ggc ttc ttg cat tct 384Lys Thr Tyr Gln Gly Ser Tyr Gly Phe Arg Leu Gly Phe Leu His Ser 115 120 125ggg aca gcc aag tct gtg act tgc acg tac tcc cct gcc ctc aac aag 432Gly Thr Ala Lys Ser Val Thr Cys Thr Tyr Ser Pro Ala Leu Asn Lys 130 135 140atg ttt tgc caa ctg gcc aag acc tgc cct gtg cag ctg tgg gtt gat 480Met Phe Cys Gln Leu Ala Lys Thr Cys Pro Val Gln Leu Trp Val Asp145 150 155 160tcc aca ccc ccg ccc ggc acc cgc gtc cgc gcc atg gcc atc tac aag 528Ser Thr Pro Pro Pro Gly Thr Arg Val Arg Ala Met Ala Ile Tyr Lys 165 170 175cag tca cag cac atg acg gag gtt gtg agg cgc tgc ccc cac cat gag 576Gln Ser Gln His Met Thr Glu Val Val Arg Arg Cys Pro His His Glu 180 185 190cgc tgc tca gat agc gat ggt ctg gcc cct cct cag cat ctt atc cga 624Arg Cys Ser Asp Ser Asp Gly Leu Ala Pro Pro Gln His Leu Ile Arg 195 200 205gtg gaa gga aat ttg cgt gtg gag tat ttg gat gac aga aac act ttt 672Val Glu Gly Asn Leu Arg Val Glu Tyr Leu Asp Asp Arg Asn Thr Phe 210 215 220cga cat agt gtg gtg gtg ccc tat gag ccg cct gag gtt ggc tct gac 720Arg His Ser Val Val Val Pro Tyr Glu Pro Pro Glu Val Gly Ser Asp225 230 235 240tgt acc acc atc cac tac aac tac atg tgt aac agt tcc tgc atg ggc 768Cys Thr Thr Ile His Tyr Asn Tyr Met Cys Asn Ser Ser Cys Met Gly 245 250 255ggc atg aac cgg agg ccc atc ctc acc atc atc aca ctg gaa gac tcc 816Gly Met Asn Arg Arg Pro Ile Leu Thr Ile Ile Thr Leu Glu Asp Ser 260 265 270agt ggt aat cta ctg gga cgg aac agc ttt gag gtg cgt gtt tgt gcc 864Ser Gly Asn Leu Leu Gly Arg Asn Ser Phe Glu Val Arg Val Cys Ala 275 280 285tgt cct ggg aga gac cgg cgc aca gag gaa gag aat ctc cgc aag aaa 912Cys Pro Gly Asg Asp Arg Thg Thr Glu Glu Glu Asn Leu Arg Lys Lys 290 295 300ggg gag cct cac cac gag ctg ccc cca ggg agc act aag cga gca ctg 960Gly Glu Pro His His Glu Leu Pro Pro Gly Ser Thr Lys Arg Ala Leu305 310 315 320ccc aac aac acc agc tcc tct ccc cag cca aag aag aaa cca ctg gat 1008Pro Asn Asn Thr Ser Ser Ser Pro Gln Pro Lys Lys Lys Pro Leu Asp 325 330 335gga gaa tat ttc acc ctt cag atc cgt ggg cgt gag cgc ttc gag atg 1056Gly Glu Tyr Phe Thr Leu Gln Ile Arg Gly Arg Glu Arg Phe Glu Met 340 345 350ttc cga gag ctg aat gag gcc ttg gaa ctc aag gat gcc cag gct ggg 1104Phe Arg Glu Leu Asn Glu Ala Leu Glu Leu Lys Asp Ala Gln Ala Gly 355 360 365aag gag cca ggg ggg agc agg gct cac tcc agc cac ctg aag tcc aaa 1152Lys Glu Pro Gly Gly Ser Arg Ala His Ser Ser His Leu Lys Ser Lys 370 375 380aag ggt cag tct acc tcc cgc cat aaa aaa ctc atg ttc aag aca gaa 1200Lys Gly Gln Ser Thr Ser Arg His Lys Lys Leu Met Phe Lys Thr Glu385 390 395 400ggg cct gac tca gac tga 1218Gly Pro Asp Ser Asp --- 405