大鼠面神经核小GTP结合蛋白TC10的表达和真核表达载体的构建

CLONING OF FULL LENGH cDNA AND EXPRESSION OF RAT SMALL GTP-BINDING PROTEIN TC10 IN FACIAL NUCLEUS INDUCED BY INJURY

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 explore the expression of small GTP-binding protein TC10 during nerve injury and construct eukaryotic expression vector carrying rat TC10 cDNA. Methods Total RNA was extracted from rat facial nucleus, and the change of rat TC10 full-length cDNA was investigated by RT-PCR. PCR product was inserted into eukaryotic expression vector pcDNA3 to construct the pcDNA 3-TC10. The plasmid was identified by restriction enzyme analysis and sequencing analysis. Results After injury, the expression of TC10 rapidly increased, Restriction enzyme assay and sequencing assay showed that the construct we got was rat TC10 full-length cDNA construct.