[发明专利]人促甲状腺素受体胞外区氨基端基因表达产物及制备方法和该产物在酶免技术中的应用

摘要

本发明公开了人促甲状腺素受体胞外区氨基端基因表达产物及制备方法和该产物在酶免技术中的应用。本发明的目的在于将TSHR－ecd cDNA分为三段(hTSHR－N端、hTSHR－C端和hTSHR－M段)分别进行、特别是对hTSHR－N端进行基因克隆、表达载体构建、E.coli原核表达，并进一步以此基因工程片段蛋白为抗原建立TRAb(包括TSAb、TSBAb)ELISA检测技术。为今后进一步TSHR抗原决定簇的研究开辟新的途径并最终转化为TSAb、TSBAb临床诊断试剂盒奠定基础。

主权项

1.一种人促甲状腺素受体胞外区氨基端基因(hTSHR-N)表达产物，其特征在于该hTSHR-N片段进行表达获重组片段蛋白(TrxFus-hTSHR-N)，其cDNA的核苷酸和氨基酸序列为：ATG GGG TGT TCG TCT CCA CCC TGC GAG TGC CAT CAG GAG GAG GAC TTC 211bpMet Gly Cys Ser Ser Pro Pro Cys Glu Cys His Gln Glu Glu Asp Phe 16AA1 7 13AGA GTC ACC TGC AAG GAT ATT CAA CGC ATC CCC AGC TTA CCG CCC AGT 259bpArg Val Thr Cys lys Asp Ile Gln Arg lle Pro Ser Leu Pro Pro Ser 32AA 19 25 31ACG CAG ACT CTG AAG CTT ATT GAG ACT CAC CTG AGA ACT ATT CCA AGT 307bpThr Gln Thr Leu lys Leu Ile Glu Thr His Leu Arg Thr Ile Pro Ser 48AA 37 43CAT GCA TTT TCT AAT CTG CCC AAT ATT TCC AGA ATC TAC GTA TCT ATA 355bpHis Ala Phe Ser Asn Leu Pro Asn Ile Ser Arg Ile Tyr Val Ser Ile 64AA49 55 61GAT GTG ACT CTG CAG CAG CTG GAA TCA CAC TCC TTC TAC AAT TTG AGT 403bpAsp Val Thr Leu Gln Gln Leu Glu Ser His Ser Phe Tyr Asn Leu Ser 80AA 67 73 79AAA GTG ACT CAC ATA GAA ATT CGG AAT ACC AGG AAC TTA ACT TAC ATA 451bplys Val Thr His Ile Glu Ile Arg Asn Thr Arg Asn Leu Thr Tyr Ile 96AA 85 91GAC CCT GAT GCC CTC AAA GAG CTC CCC CAC CTA AAG TTC CTT GGC ATT 499bpAsp Pro Asp Ala Leu lys Glu Leu Pro His Leu lys Phe Leu Gly Ile 112AA97 103 109TTC AAC ACT GGA CTT AAA ATG TTC CCT GAC CTG ACC AAA GTT TAT TCC 547bpPhe Asn Thr Gly Leu lys Met Phe Pro Asp Leu Thr lys Val Tyr Ser 128AA 115 121 127ACT GAT ATA TTC TTT ATA CTT GAA ATT ACA GAC AAC CCT TAC ATG ACG 595bpThr Asp Ile Phe Phe Ile Leu Glu Ile Thr Asp Asn Pro Tyr Met Thr 144AA 133 139TCA ATC CCT GTG AAT GCT TTT CAG GGA CTA 625bpSer Ile Pro Val Asn Ala Phe Gln Gly Leu 154AA145 151 。