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目的 :用基因工程方法获得小鼠VEGFR - 2 (Flk - 1)胞外段与配体结合区域的重组蛋白 ,并对其应用进行初步的探讨。方法 :将经PT -PCR扩增的目的序列组建到原核表达载体并诱导目的蛋白的表达 ,经纯化后的重组蛋白制备成疫苗 ,免疫兔 ,获得抗血清 ,用ELISA和Westernblot方法对抗血清进行分析。结果 :目的蛋白在原核表达系统中得到了高效的表达 ,该疫苗能刺激兔产生抗重组蛋白和天然Flk - 1的抗体。结论 :该抗血清可用于Flk - 1的免疫学检测 ,也为该疫苗用于抗肿瘤血管生成治疗奠定了基础。
OBJECTIVE: To obtain the recombinant protein of the extracellular domain of VEGFR - 2 (Flk - 1) and its ligand binding region by genetic engineering and to discuss its application. Methods: The target sequence amplified by PT-PCR was constructed into the prokaryotic expression vector and the expression of the target protein was induced. The purified recombinant protein was used to prepare the vaccine. The rabbit was immunized to obtain the antiserum. The antiserum was analyzed by ELISA and Western blot . Results: The target protein was highly expressed in prokaryotic expression system. The vaccine can stimulate rabbit to produce anti - recombinant protein and natural Flk - 1 antibody. Conclusion: The antiserum can be used in the immunological detection of Flk - 1 and lay the foundation for the anti - tumor angiogenesis therapy.