论文部分内容阅读
目的 构建含有碱性成纤维细胞生长因子 (basicfibroblastgrowthfactor,bFGF)和Math1基因的真核共表达载体 ,并用阳离子脂质体包裹将其导入大鼠耳蜗 ,观察其表达情况。方法 应用基因重组技术和限制性内切酶酶切 ,构建并鉴定PRK5 -bFGF -Math1真核共表达载体。经脂质体介导转染大鼠耳蜗后 ,用逆转录 -聚合酶链反应 (reversetranscription -polymerasechainreaction ,RT -PCR)法检测其表达情况。结果 阳性重组子经酶切鉴定含有bFGF和Math1基因 ,RT -PCR结果表明bFGF和Math1在耳蜗中均有表达。结论 本文成功地构建了含有bFGF和Math1基因的真核共表达载体 ,且在哺乳动物耳蜗中均有表达。
Objective To construct a eukaryotic co-expression vector containing basic fibroblast growth factor (bFGF) and Math1 gene, and introduce it into the cochlea of rats with cationic liposomes to observe its expression. Methods Recombinant PRK5-bFGF-Math1 eukaryotic expression vector was constructed and identified by restriction enzyme digestion and restriction endonuclease digestion. The liposome-mediated transfection of the rat cochlea, reverse transcriptase-polymerase chain reaction (reversetranscription-polymerase chain reaction (RT -PCR) method to detect the expression. Results The positive recombinant was identified by restriction enzyme digestion of bFGF and Math1, RT-PCR results showed that both bFGF and Math1 were expressed in the cochlea. Conclusion The eukaryotic co-expression vector containing bFGF and Math1 gene was successfully constructed and expressed in mammalian cochlea.