逆转录病毒载体介导人类G6PD基因在人红白血病细胞中的表达

目的：探讨逆转录病毒载体介导人类G6PD基因在人白血病细胞中的表达。方法：构建G6PD cDNA的逆转录病毒表达载体pLG6PDSN,转染包装细胞PA317，病毒上清感染人红白血病细胞K562，以PCR方法检测病毒载体是否整合于细胞基因组，定量法测定G6PD表达。t检验比较转染组与对照组间的表达差异。结果：酶切鉴定表明，G6PD cDNA准确插入pLXSN相应位点，载体构建成功。转染后PCR扩增NeoR基因，证明细胞DNA整合有逆转录病毒载体。转染组与对照组酶活性测定差异有显著性（P＜0．01）。结论：本试验所构建的重组载体pLG6PDSN为严重G6PD缺陷症的基因治疗提供了表达载体。

Expression of human G6PD gene in K562 cells mediated by retroviral vector

Objective This study aimed to investigate the feasibility of gene therapy for severe G6PD deficiency. Methods The recombinant retroviral vector bearing normal human G6PD cDNA was constructed and transferred into the erythroleukemia cell line K562. Author identified the integration of NeoR gene in the targeted cellular DNA by means of specific PCR. Quantitative method was used to measure the expression of G6PD in the targeted cells. Results Construction of the recombinant retroviral vector was successfully established. PCR indicated the integration of NeoR gene in the targeted genomic DNA of the cells. The vector was also shown to be capable of expressing the foreign gene compared to the control ( P <0.01). Conclusion The recombinant retroviral vector is competent for transferring and expressing the G6PD gene.