【作者】 郑桦锐；

【导师】 方盛国；

【作者基本信息】 浙江大学 ， 遗传学， 2002， 硕士

【摘要】 胶质细胞源神经营养因子(glial cell line-derivedneurotrophic factor，GDNF)是Lin等人于1993年从B49胶质细胞系中首次分离并纯化得到的。研究结果表明，GDNF对于神经系统具有很强的保护及恢复作用，对视网膜变性的感光细胞具有挽救作用。此外，GDNF在外周组织如肾脏的发育和精子的发生等方面，具有显著的调节功能。 本项研究经PCR引物设计后，以大熊猫和朱鹮的基因组DNA为模板，克隆了大熊猫和朱鹮的GDNF成熟蛋白基因。序列比较表明，大熊猫和朱鹮的GDNF基因与人、猕猴、大鼠、小鼠及鸡的GDNF具有非常高的同源性，分别为86.8％和75.4％、86.8％和76.6％、82.3％和68.7％、83.1％和67.9％，70.1％和94.3％。将大熊猫和朱鹮的GDNF基因片段分别克隆至pGEX—4T—3表达载体，并经IPTG诱导，表达出了预期大小的蛋白，该蛋白用Glutathione Sepharose 4BMicroSpin Column进行了纯化。更多还原

【Abstract】 Glial cell line-derived neurotrophic factor(GDNF) was initially purified and cloned from B49 glial cell line in 1993. Many studies show that GDNF has strong protection and restoration effect on rodent and primate models of neurodegenerative diseases. Recent study indicates that GDNF exerts both histologic and functional neuroprotective effects on rod photoreceptors in the rd/rd mouse. Furthermore, GDNF also play an important role in kidney development and spermatogenesis.Based on the GDNF gene sequence of human and chick, we cloned partial DNA sequences encoding mature protein of GDNF from genomic DNA of giant panda and crested ibis through PCR amplification. Sequence comparison showed the giant panda and crested ibis GDNF genes had high homology to counterparts of human, rhesus monkey, rat, mouse and chick. We constructed the pGEX-GDNF expression vecto’rs and obtained the recombinant protein of expected size after induction of IPTG. The fusion protein was purified using Glutathione Sepharose 4B affinity chromatography.更多还原