中文摘要：

目的：包装带有人U6启动子及c—Met短发夹环的重组腺相关病毒，为抑制c-Met的表达及进行肿瘤基因治疗研究奠定基础。方法：通过PCR方法将带有c-Met短发夹结构的片段构建至人U6启动子下游，将U6shMet构建至腺相关病毒载体质粒pSNAV中，将pSNAVUshMet转染BHK细胞，经G418筛选后加辅毒rHSV1-repcap包装成携带U6shMet的重组腺相关病毒。SDS-PAGE电泳分析病毒纯度，斑点杂交测定病毒滴度。结果：获得了2段带有c-Met短发夹结构的片段U6shMetl和U6shMet2，成功包装了2个带有U6shMet序列的重组腺相关病毒rAAVUshMetl和rAAVushMet2，纯化后的病毒电泳分析条带清晰、杂带少，病毒滴度均为4×10^12mg·L^-1。结论：成功构建了带有U6shMet的重组腺相关病毒rAAVUshMet，病毒纯度好、滴度高，为抑制c-Met的表达及进行肿瘤基因治疗提供了运载工具。

英文摘要：

Objective To package the recombination adeno-associated virus with human U6 promoter and c-met short hairpin in order to establish foundation for research on inhibiting the expression of c-Met and cancer gene therapy. Methods c-Met short hairpin was synthesized and linked to the down stream of U6 promoter by PCR. Adeno-associated viral vectors pSNAVUshMet （1, 2） were constructed and transfected into BHK cells. Positive cell clones were selected by G418. Adeno-associated virus with U6shMet were packaged by adding rHSVl-repcap. The virus purity was analysed by SDS-PAGE and titer was assayed by dot blot. Results Two fragments （U6shMetl and U6shMet2） with c-Met short hairpin were obtained and two recombination adeno-associated virus （rAAVUshMetl and rAAVUshMet2） with U6shMet were packaged successfully. SDS-PAGE analysis showed that the purified virus appeared clear characterized bands. The virus titer was 4 × 10^12 mg · L^-1. Conclusion The recombination adeno-associated virus （rAAVUshMetl and rAAVUshMet2） with U6shMet are constructed successfully. The virus have high titer and good purity and could act as the effective vehicle of inhibiting the expression of c-Met and cancer gene therapy.