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目的 :构建人整合素 β3亚基真核表达载体 ,并探讨如何使人整合素αIIbβ3在CHO细胞表面有效表达 ,为进一步研究整合素β3亚基作为汉坦病毒 (hantavirus,HV)受体的特异性奠定基础。方法 :构建编码人整合素 β3真核表达载体 pcDNA3.1 β3。将其与编码人整合素αIIb亚基真核表达载体 ,分别及共转染至中国仓鼠卵巢 (Chinahamsterovary ,CHO)细胞中进行表达。采用间接免疫荧光法 (IFA)检测外源基因的表达。结果 :共转染组目的蛋白呈高效的细胞膜表达。pcDNA3.1 β3单独转染组 β3亚基在细胞膜的表达较共转染组弱 ;而pBJ1 αIIb单独转染组则未见有效的细胞膜表达。结论 :人整合素αIIbβ3在CHO细胞表面的有效表达需要两个亚基共同参与。
OBJECTIVE: To construct eukaryotic expression vector of human integrin β3 subunit and to explore how to make human integrin αIIbβ3 efficiently expressed on CHO cell surface. To further investigate the specificity of integrin β3 subunit as a receptor of hantavirus (HV) Sex laid the foundation. Methods: Construction of human integrin β3 eukaryotic expression vector pcDNA3.1 β3. The recombinant eukaryotic expression vector of human integrin αIIb subunit was co-transfected into Chinese hamster ovary (CHO) cells for expression. Indirect immunofluorescence (IFA) was used to detect the expression of foreign genes. Results: The target protein in the co-transfection group was highly efficient in cell membrane expression. The expression of β3 subunit of pcDNA3.1β3 alone transfected group was weaker than that of cotransfection group. However, pBJ1 αIIb alone did not show any significant cell membrane expression. Conclusion: The effective expression of human integrin αIIbβ3 on the surface of CHO cells requires the participation of two subunits.