摘要
本文通过PCR技术,直接从大熊猫基因组DNA上克隆得到其神经营养素-4的成熟肽编码序列,通过序列分析发现,该基因在进化上具有较高的保守性。将神经营养素-4成熟肽完整编码序列克隆至pGEX-4T-3表达载体,并经IPTG诱导在大肠杆菌中进行原核生物表达,获得了大熊猫重组蛋白神经营养素-4。重组表达蛋白经纯化后,进行大鼠肾上腺嗜铬瘤细胞神经营养因子的活性鉴定,发现其能够诱导神经细胞分化产生突触,具有预期的生物学活性。对大熊猫神经营养素-4的基因工程研究,为大熊猫癫痫的基因治疗奠定了基础。
The neurotrophin-4 (NT-4) gene plays an important role in the protection of the damages resulted from epilepsy, which is an important cause of death in the pandas. The genomic DNA coding for the NT-4 gene of the giant panda was cloned into expression vector pGEX-4T-3 under the control of a tac promoter, and expressed in Escherichia coli. After purification, the recombinant NT-4 of the panda was found to be biologically active in the PC12 neurite outgrowth assay. We suggest that this technique may offer a feasible means of gene therapy for the giant pandas suffering from epilepsy.
出处
《兽类学报》
CAS
CSCD
北大核心
2004年第1期13-18,共6页
Acta Theriologica Sinica
基金
StateKeyBasicResearchandDevelopmentPlanofP R ofChina (G2 0 0 0 0 4680 6)
关键词
癫痫
神经营养素-4
基因表达
纯化
大熊猫
基因治疗
Epilepsy
Neurotrophin-4
Gene expression
Purification
Giant panda(Ailuropoda melanoleuca)
