中国肿瘤生物治疗杂志2001，Vol.8Issue(1)：52-54,3.
D-氨基酸氧化酶基因重组逆转录病毒载体的构建及表达
Construction and Expression of Recombinant Retroviral Vector Containing D-Amino Acid Oxidase cDNA
摘要
Abstract
Objective: To construct retroviral vector pLDAAOSN and observethe function of DAAO gene. Methods: With recombinant DNA technology, DAAO cDNA was cloned into retroviral vector (pLDAAOSN). The vector was transfected into ΦXNA cells by CaPO4 method, and the DAAO cDNA was transferred by recombinant retroviral vector into leukemia cell line K562. The positive clones were obtained after G418 selection and termed KDAAO. PCR and in situ hybridization were used to identify the integration and expression of DAAO gene in KDAAO. In order to observe the function of DAAO, KDAAO was treated with 50 mm/L D-Ala. Results: Results of plasmid pLDAAOSN digested with KpnⅠ and the sequence determination confirmed pLDAAOSN contains full-length DAAO cDNA. Infectious titer generated by the packaging cells was 5.2×106 CFU/ml. PCR and in situ hybridization analysis showed integration of DAAO gene in KDAAO and expression of DAAO gene at mRNA level. Preliminary observation suggested that D-Ala could effectively kill KDAAO. Conclusion: Retroviral vector pLDAAOSN may be useful for futher study of gene therapy in cancer.关键词
逆转录病毒载体/DAAO基因/白血病细胞系分类
医药卫生引用本文复制引用
翟勇平,王健民,周虹,张雨生..D-氨基酸氧化酶基因重组逆转录病毒载体的构建及表达[J].中国肿瘤生物治疗杂志,2001,8(1):52-54,3.基金项目
本课题由国家自然科学基金重点项目(39870710)资助 （39870710）
