人的细胞间粘附分子－1膜外I－Ⅲ功能区基因表达产物及其制备方法和该产物在放免技术中的应用

摘要

本发明公开了一种人的细胞间粘附分子-1膜外I-III功能区基因表达产物及其制备方法和该产物在放免技术中的应用，属于基因工程技术领域。将ICAM-1膜外I-III功能区编码基因重组到表达型质粒pET42a中，并将其转化入原核表达菌中，纯化获得表达产物GST-ICAM-1；以此作为抗原，免疫家兔后获得兔抗人的多克隆抗体。这样制备的人的细胞间粘附分子-1膜外I-III功能区基因在原核系统的中表达产物较真核系操作简便，更易于纯化，价格便宜，产率高。在国内外率先研制出人血清sICAM-1放射免疫分析技术，并初步应用于临床检测，对于判断Graves’病的疗效、停药和复发具有重要意义。

主权项

1.一种人的细胞间粘附分子-1膜外I-III功能区基因表达产物，其特征在于该表达产物是一种具有人的细胞间粘附分子-1蛋白抗原性的重组蛋白，其cDNA的核苷酸和氨基酸序列为：CAG ACA TCT GTG TCC CCC TCA AAA GTC ATC CTG CCC CGG GGA GGC TCC GTG 189Gln Thr Ser Val Ser Pro Ser Lys Val Ile Leu Pro Arg Gly Gly Ser Val 17CTG GTG ACA TGC AGC ACC TCC TGT GAC CAG CCC AAG TTG TTG GGC ATA GAG 240Leu Val Thr Cys Ser Thr Ser Cys Asp Gln Pro Lys Leu Leu Gly Ile Glu 34ACC CCG TTG CCT AAA AAG GAG TTG CTC CTG CCT GGG AAC AAC CGG AAG GTG 291Thr Pro Leu Pro Lys Lys Glu Leu Leu Leu Pro Gly Asn Asn Arg Lys Val 51TAT GAA CTG AGC AAT GTG CAA GAA GAT AGC CAA CCA ATG TGC TAT TCA AAC 342Tyr Glu Leu Ser Asn Val Gln Glu Asp Ser Gln Pro Met Cys Tyr Ser Asn 68TGC CCT GAT GGG CAG TCA ACA GCT AAA ACC TTC CTC ACC GTG TAC TGG ACT 393Cys Pro Asp Gly Gln Ser Thr Ala Lys Thr Phe Leu Thr Val Tyr Trp Thr 85CCA GAA CGG GTG GAA CTG GCA CCC CTC CCC TCT TGG CAG CCA GTG GGC AAG 444Pro Glu Arg Val Glu Leu Ala Pro Leu Pro ser Trp Gln Pro Val Gly Lys 102AAC CTT ACC CTA CGC TGC CAG GTG GAG GGT GGG GCA CCC CGG GCC AAC CTC 495Asn Leu Thr Leu Arg Cys Gln Val Glu Gly Gly Ala Pro Arg Ala Asn Leu 119ACC GTG GTG CTG CTC CGT GGG GAG AAG GAG CTG AAA CGG GAG CCA GCT GTG 546Thr Val Val Leu Leu Arg Gly Glu Lys Glu Leu Lys Arg Glu Pro Ala Val 136GGG GAG CCC GCT GAG GTC ACG ACC ACG GTG CTG GTG AGG AGA GAT CAC CAT 597Gly Glu Pro Ala Glu Val Thr Thr Thr Val Leu Val Arg Ar Aspg His His 153GGA GCC AAT TTC TCG TGC CGC ACT GAA CTG GAC CTG CGG CCC CAA GGG CTG 648Gly Ala Asn Phe Ser Cys Arg Thr Glu Leu Asp Leu Arg Pro Gln Gly Leu 170GAG CTG TTT GAG AAC ACC TCG GCC CCC TAC CAG CTC CAG ACC TTT GTC CTG 699Glu Leu Phe Glu Asn Thr Ser Ala Pro Tyr Gln Leu Gln Thr Phe Val Leu 187CCA GCG ACT CCC CCA CAA CTT GTC AGC CCC CGG GTC CTA GAG GTG GAC ACG 750Pro Ala Thr Pro Pro Gln Leu Val Ser Pro Arg Val Leu Glu Val Asp Thr 204CAG GGG ACC GTG GTC TGT TCC CTG GAC GGG CTG TTC CCA GTC TCG GAG GCC 801Gln Gly Thr Val Val Cys Ser Leu Asp Gly Leu Phe Pro Val Ser Glu Ala 221CAG GTC CAC CTG GCA CTG GGG GAC CAG AGG TTG AAC CCC ACA GTC ACC TAT 852Gln Val His Leu Ala Leu Gly Asp Gln Arg Leu Asn Pro Thr Va Thr Tyrl 238GGC AAC GAC TCC TTC TCG GCC AAG GCC TCA GTC AGT GTG ACC GCA GAG GAC 903Gly Asn Asp Ser Phe Ser Ala Lys Ala Ser Val Ser Val Thr Ala Glu Asp 255GAG GGC ACC CAG CGG CTG ACG TGT GCA GTA ATA CTG GGG AAC CAG AGC CAG 954Glu Gly Thr Gln Arg Leu Thr Cys Ala Val Ile Leu Gly Asn Gln Ser Gln 272GAG ACA CTG CAG ACA GTG ACC ATC TAC AGC TTT CCG 990Glu Thr Leu Gln Thr Val Thr Ile Tyr Ser Phe Pro 284