重组人干扰素α2a和胸腺肽α1融合蛋白的分子设计及其基因表达产物的鉴定

人干扰素α_2a（IFNα_2a）和胸腺肽α₁（THYα₁）联合使用效果远大于各自单独使用的效果，本文通过DNA重组技术将两者构建成一个融合蛋白药物。首先，依据IFNα_2a和THYα₁的分子结构，设计了一个连接肽，将IFNα_2a和THYα₁相连，并分别位于该融合蛋白的N端和C端。在此基础上，构建出表达载体pCJ101质粒，获得的阳性克隆307在2×YT培养基小试中，产物变性复性后经DEAE-Sepharose柱层析纯化后，在SDS-PAGE上得到相对分子质量为23000的谱带。经鉴定，融合蛋白中干扰素α_2a的比酶活性为1.34mol/s·g；检测胸腺肽α₁的活性，显示活性E玫瑰花结形成率达到20%。结果表明，融合蛋白中两种药物蛋白均较好地保持各自的生物功能，为继续研究奠定了良好基础。

Molecular design and characterization of recombinant fusion protein of human IFNα2a-THYα1

The combinated IFNα_2a and THYα₁ is more powerful than the individual of both IFNα_2a and THYα₁ medicines during treating human cancers and some virus infection diseases, and we considered to construct a fusion protein molecule containing both IFNα_2a and THYα₁ by DNA recombination. First of all, based on the three dimension structure of both molecules, a linker with 12 amino acid length was designed to connect to IFNα_2a C terminal and THYα₁ N terminal. After that, an expressed vector pCJ101 containing the fusion gene was obtained and transformed into host cell E.coli DH5α. The main expressed protein band on SDS-PAGE possessed 23000 in molecular weight. The biofunction test of the fusion protein showed the activity of IFNα_2a reached 1.834×10^-2mol/（s·mg） and THYα₁ reached 20%(E-rosette formation rate), respectively.