【作者】 白忠彬；

【导师】 金成汉；

【作者基本信息】 延边大学 ， 基础兽医学， 2002， 硕士

【摘要】 生长激素释放因子(Growth Hormone Releasing Factor，GRF)是由动物下丘脑合成并分泌的一种多肽激素，能特异性的诱导生长激素的合成与分泌，提高体内生长激素的水平。本实验是将含GRF重组质粒的JM109菌株大量培养，用碱裂变法提取质粒，用DNA Caculator定量；制备含GRF的原生质体，经1％的戊二醛处理后注射于家兔肌肉，在注射部位给予电刺激，提取总RNA，用RT-PCR检测GRF在肌肉中的表达；用ELISA法定性检测GRF在肌肉中蛋白质水平的表达；将该原生质体注射于小鼠肌肉中，定期提取DNA，初步探讨原生质介导的外源性GRF在小鼠肌肉中的表达时间。 研究结果表明，以原生质体介导的外源性GRF基因可以在家兔和小鼠肌肉中表达，通过PCR、RT-PCR和ELISA都可以检测到表达产物。电击可以促进原生质体与肌肉细胞的融合，但100V，50毫秒的低压、短脉冲电击对GRF在动物体内的表达量上没有被观察到显著差异。通过提取注射部位DNA后进行PCR鉴定，可以确认原生质体介导的GRF基因至少可以在受试动物体内滞留28天以上，为GRF的长期表达提供了条件。增重实验表明，原生质体介导的外源性GRF基因对家兔的增重有明显促进作用。更多还原

【Abstract】 Growth Hormone Releasing Factor is Polypeptide hormone releasing by animal hypothaiami. It can induction synthesis and excretion of growth hormone which rise growth hormone level of animal. We large-scale cultured JM-109 strain recombinant plasmid of GRF. Then the plasmid was extracted from them and determined by DNA calculator. The protoplast that contain growth hormone releasing factor injected into rabbit muscle and mouse muscle after it were treated by 1% Glutaral,pay it to electric stimulation in muscle of injection site and extract omni-RNA from injection site of rabbit muscle,expression of GRF were detected by reverse transcription and PCR;ratio of expression of GRF were detected by ELISA. Extract DNA form injection site of mouse muscle to research time of expression .The result of investigation showing:extro growth hormone releasing factor mediated by protoplast expressed in rabbit muscle and mouse muscle,products of expression were detected by PCR RT-PCR and ELISA. Electroporation will acceleration cellular confluence of protoplast and muscles,but the significant changeof the ratio of expression and the time of expression was not observed form exponent treat with low pressure and short impulse of 100V,50 millisecond. Extro growth hormone releasing factor mediated by protoplast could more than 28 days retention in rabbit and mouse muscle that is condition of long-time expression of GRF.更多还原