[发明专利]双重靶效应基因嵌合重组体及其构建方法和应用

摘要

本发明提供了一种双重靶效应基因嵌合重组体及其构建方法和应用。用特异性引物以SDF－54/R/pET－30a(+)为模板扩增SDF－54/R，并将其直接插入T－A克隆载体pMD18－T，然后在其3’－端顺次加入人类IgG1抗体重链的铰链区编码序列和Decorin成熟肽编码序列，构建成SDF－54/R－IgG1HCHR－Decorin/pMD18－T嵌合重组体。SDF－54/R－IgG1HCHR－Decorin保留对CXCR4的亲和力而失去其内在活性，并能抑制野生型SDF－1α对乳腺癌细胞的趋化迁移活性、抑制乳腺癌细胞的生长，其可作为CXCR4的新型拮抗剂—同时抑制CXCR4阳性肿瘤细胞的生长和转移，具有双重靶效应。

主权项

1、一种双重靶效应基因嵌合重组体，其特征在于，所述双重靶效应基因嵌合重组体核酸编码序列为：aagcccgtcagcctgagctacagatgcccatgccgattcttcgaaagccatgttgccagagccaacgtcaagcatctcaaaattctcaacactccaaactgtgcccttcagattgtagcccggctgaagaacaacaacagacaagtgtgcattgacccgaagctcgagCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAggatccgatgaggcttctgggataggcccagaagttcctgatgaccgcgacttcgagccctccctaggcccagtgtgccccttccgctgtcaatgccatcttcgagtggtccagtgttctgatttgggtctggacaaagtgccaaaggatcttccccctgacacaactctgctagacctgcaaaacaacaaaataaccgaaatcaaagatggagactttaagaacctgaagaaccttcacgcattgattcttgtcaacaataaaattagcaaagttagtcctggagcatttacacctttggtgaagttggaacgactttatctgtccaagaatcagctgaaggaattgccagaaaaaatgcccaaaactcttcaggagctgcgtgcccatgagaatgagatcaccaaagtgcgaaaagttactttcaatggactgaaccagatgattgtcatagaactgggcaccaatccgctgaagagctcaggaattgaaaatggggctttccagggaatgaagaagctctcctacatccgcattgctgataccaatatcaccagcattcctcaaggtcttcctccttcccttacggaattacatcttgatggcaacaaaatcagcagagttgatgcagctagcctgaaaggactgaataatttggctaagttgggattgagtttcaacagcatctctgctgttgacaatggctctctggccaacacgcctcatctgagggagcttcacttggacaacaacaagcttaccagagtacctggtgggctggcagagcataagtacatccaggttgtctaccttcataacaacaatatctctgtagttggatcaagtgacttctgcccacctggacacaacaccaaaaaggcttcttattcgggtgtgagtcttttcagcaacccggtccagtactgggagatacagccatccaccttcagatgtgtctacgtgcgctctgccattcaactcggaaactataag；所述双重靶效应基因嵌合重组体氨基酸序列为：Lys Pro Val Ser Leu Ser Tyr Arg Cys Pro Cys Arg Phe Phe Glu SerHis Val Ala Arg Ala Asn Val Lys His Leu Lys Ile Leu Asn Thr ProAsn Cys Ala Leu Gln Ile Val Ala Arg Leu Lys Asn Asn Asn Arg GlnVal Cys Ile Asp Pro Lys Leu Glu Pro Lys Ser Cys Asp Lys Thr HisThr Cys Pro Pro Cys Pro Gly Ser Asp Glu Ala Ser Gly Ile Gly ProGlu Val Pro Asp Asp Arg Asp Phe Glu Pro Ser Leu Gly Pro Val CysPro Phe Arg Cys Gln Cys His Leu Arg Val Val Gln Cys Ser Asp LeuGly Leu Asp Lys Val Pro Lys Asp Leu Pro Pro Asp Thr Thr Leu LeuAsp Leu Gln Asn Asn Lys Ile Thr Glu Ile Lys Asp Gly Asp Phe LysAsn Leu Lys Asn Leu His Ala Leu Ile Leu Val Asn Asn Lys Ile SerLys Val Ser Pro Gly Ala Phe Thr Pro Leu Val Lys Leu Glu Arg LeuIyr Leu Ser Lys Asn Gln Leu Lys Glu Leu Pro Glu Lys Met Pro LysThr Leu Gln Glu Leu Arg Ala His Glu Asn Glu Ile Thr Lys Val ArgLys Val Thr Phe Asn Gly Leu Asn Gln Met Ile Val Ile Glu Leu GlyThr Asn Pro Leu Lys Ser Ser Gly Ile Glu Asn Gly Ala Phe Gln GlyMet Lys Lys Leu Ser Tyr Ile Arg Ile Ala Asp Thr Asn Ile Thr SerIle Pro Gln Gly Leu Pro Pro Ser Leu Thr Glu Leu His Leu Asp GlyAsn Lys Ile Ser Arg Val Asp Ala Ala Ser Leu Lys Gly Leu Asn AsnLeu Ala Lys Leu Gly Leu Ser Phe Asn Ser Ile Ser Ala Val Asp AsnGly Ser Leu Ala Asn Thr Pro His Leu Arg Glu Leu His Leu Asp AsnAsn Lys Leu Thr Arg Val Pro Gly Gly Leu Ala Glu His Lys Tyr IleGln Val Val Tyr Leu His Asn Asn Asn Ile Ser Val Val Gly Ser SerAsp Phe Cys Pro Pro Gly His Asn Thr Lys Lys Ala Ser Tyr Ser GlyVal Ser Leu Phe Ser Asn Pro Val Gln Tyr Trp Glu Ile Gln Pro SerThr Phe Arg Cys Val Tyr Val Arg Ser Ala Ile Gln Leu Gly Asn TyrLys。