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目的:构建人肝细胞癌特异性抗原HCA661的DNA疫苗。方法:通过PCR从SMMC7721中获得编码肝细胞癌特异抗原HCA661的基因组DNA序列,插入pGEM-T easy载体后测序,并将目的基因定向亚克隆进入真核表达载体pCI-neo,以PCR和酶切方法鉴定构建的重组真核表达质粒pCI-neo-HCA661。结果:PCR产物为1 040 bp,测序结果与Gene Bank登记完全相同;PCR和酶切结果表明,构建的重组真核表达质粒中含有正确编码的目的基因片段。结论:成功地构建了含有HCA661基因的DNA疫苗。
Objective: To construct a DNA vaccine of human hepatocellular carcinoma-specific antigen HCA661. Methods: Genomic DNA sequence encoding hepatocellular carcinoma-specific antigen HCA661 was obtained from SMMC7721 by PCR, inserted into pGEM-T easy vector and sequenced. The target gene was subcloned into eukaryotic expression vector pCI-neo. Methods The constructed recombinant eukaryotic expression plasmid pCI-neo-HCA661 was identified. Results: The PCR product was 1 040 bp. The sequencing result was exactly the same as that of Gene Bank. The results of PCR and restriction enzyme digestion showed that the constructed recombinant eukaryotic expression plasmid contained the correct segment of the target gene. Conclusion: The DNA vaccine containing HCA661 gene was successfully constructed.