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目的 分析大鼠小GTP结合蛋白TC10基因在大鼠面神经损伤早期的表达和作用 ,构建大鼠TC10的真核表达载体。 方法 由大鼠损伤的面神经核中提取组织总RNA ,利用逆转录聚合酶链反应技术观察面神经损伤后面神经核团TC10的表达变化 ,将获得的基因片段酶切后插入pcDNA3真核表达载体中 ,利用酶切电泳和核苷酸序列分析鉴定。 结果 逆转录聚合酶链反应显示在正常情况下 ,面神经核团有TC10的表达 ,切断后 6h即出现增加 ,2 4h达到最大值。TC10真核表达质粒酶切电泳及序列测定证明 ,所获得的基因片段为大鼠TC10全长基因cDNA序列。 结论 面神经切断后 ,TC10在面神经核团出现急剧增加 ,可能参与了神经元早期损伤反应的信号转录。本实验首次从大鼠面神经核团中获得大鼠TC10的cDNA ,并构建了TC10真核表达质粒。为研究TC10的表达及作用提供了必要条件。
Objective To analyze the expression of rat small GTP-binding protein TC10 gene in the early stage of facial nerve injury in rats and construct the eukaryotic expression vector of rat TC10. Methods Tissue RNA was extracted from the facial nerve nucleus of rats and the expression of TC10 in the facial nucleus after facial nerve injury was observed by reverse transcription polymerase chain reaction. The obtained gene fragment was digested and inserted into pcDNA3 eukaryotic expression vector, Identification by enzyme-cut electrophoresis and nucleotide sequence analysis. Results Reverse transcriptase-polymerase chain reaction showed that under normal conditions, the expression of TC10 in the facial nucleus increased after 6 hours and reached the maximum at 24 hours. TC10 eukaryotic expression plasmid digestion electrophoresis and sequencing proved that the obtained gene fragment is the full-length cDNA sequence of rat TC10. Conclusions After facial nerve transection, TC10 dramatically increases in the facial nucleus, which may be involved in the signal transduction of neuronal damage in early stage. In this study, the cDNA of rat TC10 was obtained from rat facial nerve nuclei for the first time, and a TC10 eukaryotic expression plasmid was constructed. To study the expression of TC10 and the role provided the necessary conditions.