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为了探索极低密度脂蛋白受体配体结合域中 8个重复序列在结合脂蛋白中的作用 ,利用基因缺失诱变的方法 ,构建了不同重复序列缺失的极低密度脂蛋白受体缺失突变真核表达载体 ,并将各重组体转移至体外培养的中国仓鼠卵巢细胞。通过逆转录—聚合酶链反应检测到外源基因的表达。转染细胞与DiI标记的 β 极低密度脂蛋白结合实验发现 :重复序列 1,2缺失的极低密度脂蛋白受体结合 β 极低密度脂蛋白的能力明显降低。为进一步研究极低密度脂蛋白受体结构与功能的关系奠定了基础。
In order to explore the role of eight repeats in the binding domain of very low density lipoprotein receptor ligands in the binding of lipoproteins, a very low density lipoprotein receptor deletion mutation with different repeat deletions was constructed by gene deletion mutagenesis Eukaryotic expression vector, and each recombinant was transferred to in vitro cultured Chinese hamster ovary cells. Exogenous gene expression was detected by reverse transcription-polymerase chain reaction. Binding of transfected cells to DiI-labeled beta-very-low-density lipoprotein revealed that the ability of very-low-density lipoprotein receptors with repetitive sequence 1, 2 deletion to bind beta-very low density lipoprotein was significantly reduced. Which laid the foundation for further study on the relationship between the structure and function of very low density lipoprotein receptor.