CAJ | 학술논문

采用基因克隆的方法构建人白细胞介素(hIL-24)基因的原核表达载体pET-28a(+)-hIL24,在大肠杆菌中诱导表达重组蛋白rhIL-24,SDS-PAGE和Western blotting分析蛋白表达,探讨其最佳诱导表达条件.结果表明:hIL-24基因的表达产物分子质量约25 ku,且以包涵体的形式存在于超声裂解后的沉淀中,重组蛋白rhIL-24可特异性被鼠抗人IL-24单克隆抗体识别.当IPTG终浓度为0.5 mmol·L~(-1)、37℃诱导8 h时,其表达量最高.hIL-24基因在该原核表达系统中的成功表达,为其生物学功能的研究奠定了基础.
채용기인극륭적방법구건인백세포개소(hIL-24)기인적원핵표체재체pET-28a(+)-hIL24,재대장간균중유도표체중조단백rhIL-24,SDS-PAGE화Western blotting분석단백표체,탐토기최가유도표체조건.결과표명:hIL-24기인적표체산물분자질량약25 ku,차이포함체적형식존재우초성렬해후적침정중,중조단백rhIL-24가특이성피서항인IL-24단극륭항체식별.당IPTG종농도위0.5 mmol·L~(-1)、37℃유도8 h시,기표체량최고.hIL-24기인재해원핵표체계통중적성공표체,위기생물학공능적연구전정료기출.
In this study,a recombinant expression vector of human interleukin-24(hIL-24)gene,named pET-28a(+)-hIL24,was constructed by gene cloning.The recombinant protein hIL-24 was expressed in E. coil and analyzed by SDS-PAGE and Western blotting.The proper inducing conditions of its expression were explored.The results showed that the recombinant protein was about 25 ku and mainly existed in the precipitation after sonication.Western blotting proved that the recombinant protein IL-24 was recognized by mouse anti-human IL-24.SDS-PAGE analysis indicated that the best expression condition was 0.5 mmol·L~(-1) IPTG for 8 h at 37℃.The successful expression of hIL-24 in the prokaryotic expression system is helpful for the further study of its biological function.