CAJ | 학술논문

目的：原核表达脑源性神经营养因子前体（ proBDNF ）基因重组蛋白，并检测其生物学活性。方法通过PCR方法扩增大鼠点突变型基因的脑源性神经营养因子前体的cDNA，克隆到质粒pET-28a中，在大肠杆菌BL21中表达，其表达产物经Western印迹方法鉴别，用DAPI法检测其对PC12细胞凋亡的影响。结果重组基因的扩增产物在大肠杆菌中获得表达，表达产物符合目的蛋白分子量并能够与特异性抗体反应。原核表达的proBDNF基因重组蛋白能够诱导PC12细胞的凋亡。结论成功地在大肠杆菌BL21中表达了proBDNF基因重组蛋白，该蛋白能够对促进PC12细胞的凋亡，具有生物毒性。
목적：원핵표체뇌원성신경영양인자전체（ proBDNF ）기인중조단백，병검측기생물학활성。방법통과PCR방법확증대서점돌변형기인적뇌원성신경영양인자전체적cDNA，극륭도질립pET-28a중，재대장간균BL21중표체，기표체산물경Western인적방법감별，용DAPI법검측기대PC12세포조망적영향。결과중조기인적확증산물재대장간균중획득표체，표체산물부합목적단백분자량병능구여특이성항체반응。원핵표체적proBDNF기인중조단백능구유도PC12세포적조망。결론성공지재대장간균BL21중표체료proBDNF기인중조단백，해단백능구대촉진PC12세포적조망，구유생물독성。
Objective To generate the gene recombinant protein of brain-derived neurotrophic factor precursor ( proBDNF ) in prokaryotic cells and investigate its biological activity .Methods Rat-derived cDNA of proBDNF with point mutation was amplified by polymerase chain reaction ( PCR ) and cloned into plasmid pET-28a for expression in E.coli BL21.Western blot was used to identify the product and DAPI performed to test its effect on apoptosis of PC 12 cells.Results PCR product of recombinant gene was successfully expressed in E .coli.And the product agreed with the target protein in molecular weight and showed reactivity with its specific antibody .Apoptosis of PC12 cells was induced by a certain concentration of recombinant proBDNF .Conclusion The prokaryotic expression vector has been successfully constructed for recombinant gene of proBDNF .And the product has biological toxicity and it may induce the apoptosis of PC12 cells.