P—选择素凝集样区和绿色荧光蛋白融合基因的构建

目的 构建P-选择素凝集样区与绿色荧光蛋白融合基因（pEGFP-N1/L），为进一步明确P-选择素不同表位与功能间的关系，为阐明P-选择素生理病理意义奠定基础。方法 应用PCR技术扩增质粒pCDM1/cDNA凝集素校区，用T4连接酶将其插入载体pEGFP-N1；经酶切和测序对插入片段进行分析、鉴定。结果 通过酶切和序列分析证实插入片段序列正确。结论 应用基因工程技术构建pEGFP-N1/L融合基因成功，为绿色荧光蛋白作为主生物标记分子来研究P-选择素分子的构效关系打下良好的基础。

Construction of P- selectin lectin- like domain fused with green fluorescent protein gene

Objective To construct a fused gene of P selectin lectin like domain and green fluorescent protein(pEGFP N1/L) to provide the scientific basis to clarify the pathological physiology sense of P selectin. It also has laid a foundation to analyze the relationship between molecular structure and function. Methods The lectin like region fragment of P selectin obtained by PCR was insert into pEGFP N1. The fusion gene was verified by enzyme digestive analysis and sequencing. Results A positive band was formed when product of PCR electrophoresis on agarose gel and insert fragment was confirmed correctly by enzyme digestion analysis and sequencing. Conclusion The fusion gene pEGFP N1/L was constructed successfully by genetic engineer technique. It has laid a foundation to study P selectin's structure function relationship by GFP used as biology marker molecular.