P-选择素凝集素样结构域和红色荧光蛋白融合基因的构建

Construction of fused gene of P-selectin lectin-like domain and red fluorescent protein

目的 构建P 选择素凝集素样结构域与红色荧光蛋白融合基因 (pDsRed1 N1/L) ,以进一步明确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凝集素样区 ,用T4连接酶将其插入载体pDsRed1 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确定重 ,组pDsRed1/N1的片段为目的基因的核苷酸序列。 结论 应用基因工程技术构建 pDsRed1 N1/L融合基因成功 ,为红色荧光蛋白作为生物标记分子来研究P 选择素分子的构效关系打下基础。

Objective To construct a fused gene of P-selectin lectin-like domain and red fluorescent protein(pDsRed1-N1/L) to provide the scientific basis for clarifying the pathophysiological significance of P-selectin and analyzing the relationship between molecular structure and function.Methods The lectin-like region fragment of P-selectin obtained by PCR was inserted into pDsRed1-N1/L.The fusion gene was verified by enzyme digestive analysis and sequencing.Results The sequence of isolated lectin domain of P-selectin is same as reported.Conclusion The success of constructing pDsRed1-N1/L created a way for study of structure-function relationship of P-selectin by red fluorescent protein used as biologically labelled molecular.