中文摘要：

目的构建mCD99L2（mouse CD99 antigen-like2）基因RNA干扰慢病毒表达载体，检测其对293FT细胞的感染效率。方法应用基因工程技术，首先设计并合成4对siRNA序列，退火、酶切并连接于慢病毒表达载体SDl259．构建携带针对目的基因mCD99L2的siRNA慢病毒穿梭质粒表达载体（其中包括1条阴性对照序列）；然后使用慢病毒包装质粒混合物和构建好的慢病毒穿梭质粒共转染293FT细胞，转染48h后收集上清离心过滤，浓缩病毒，利用绿色荧光蛋白作为报告基因，对病毒滴度和感染效率进行检测。结果构建3个携带针对目的基因mCD99L2的siRNA慢病毒穿梭质粒表达载体和1个阴性对照质粒，经测序鉴定正确；共转染293FT细胞包装病毒并浓缩后滴度达1×10^7／ml．适合感染目的细胞。结论应用基因工程技术成功构建了mCD99L2基因RNA干扰慢病毒表达载体，为进一步构建类人霍奇金淋巴瘤可视化细胞模型及动物模型奠定了基础。

英文摘要：

Objective To construct a recombinant lentivirus harboring RNA interference sequence targeting mouse CD99 antigen-like 2 （mCD99L2） gene and observe its infection efficiency of 293FT cells. Methods Four pairs of small interfering RNAs （siRNAs） targeting mCD99L2 cDNA were designed, synthesized and linked to the lentivirus vector SD1259 to construct the lentivirus shuttle plasmids. After sequencing, the 4 lentivirus shuttle plasmids were transfected into 293FT cells in the presence of packaging plasmids. Forty-eight hours later, the supernatant was collected and the titer and infection efficiency of the recombinant lentivirus were determined according to the expression of the reporter gene enhanced green fluorescent protein （EGFP） under fluorescent microscope. Results DNA sequencing demonstrated that mCD99L2 siRNAs were successfully cloned to the lentiviral vector SD1259. The titer of concentrated virus was 1×10^7/ml in the supernatant of the infected cells. Conclusion The recombinant lentivirus containing siRNA targeting mCD99L2 gene has been successfully constructed, which provide the basis for future establishment of visualized cell model and animal model of Hodgkin＇s lymphoma.