狼疮鼠淋巴细胞IgG2aVH区反义RNA重组体的构建及鉴定

目的 :构建系统性红斑狼疮 (SLE)鼠模型 (NZW×NZB)F1淋巴细胞IgG2aVH 区反义RNA表达重组体 ,为SLE基因治疗提供基础。方法 :采用逆转录—PCR技术克隆狼疮鼠淋巴细胞IgG2aVH 区cDNA ,经T/A克隆再定向插入表达载体pcDNA3.1,并经酶切电泳 ,PCR ,DNA测序鉴定分析。结果 :经鉴定 ,证明成功构建了表达狼疮鼠淋巴细胞IgG2aVH 区的重组体。结论 :BWF1和鼠淋巴细胞中存在IgG2aVH 区 ,其表达型重组体可用于IgG2aVH 的大量复制。

Construction of recombinant antisense RNA IgG2aVH region of murine systemic lupus erythematous

Objective To construct recombinant plasmid of IgG2aV H region antisense RNA of systemic lupus erythematous. Methods Total RNA was isolated from spleen cell of BWF1 mice. Using spleen cell total RNA as the template, We designed specific primers from A6.1 region sequences, and amplified the IgG2aV H region 375 bp DNA fragments by RT PCR. The IgG2aV H cDNA was cloned by T/A and inserted into pcDNA 3.1 plasmid of vector. Results The IgG2aV H antisense RNA plasmid expressing recombinant was identified by restriction enzyme digestion and DNA sequence analysis. Conclusion There is IgG2aV H gene in F1 mice spleen cell; the IgG2aV H antisense RNA expressing recombinants is constructed successfully.