CAJ | 학술논문

目的通过昆虫杆状病毒表达系统的优势，表达出具有生物学活性的神经营养素-4(hNT-4)蛋白。方法通过PCR方法获得hNT-4成熟肽的基因，将其连接到昆虫表达载体pAcGP67B上，在昆虫细胞Sf9中进行表达。对表达产物进行免疫原性和生物学活性测定。结果获得hNT-4蛋白的表达产物，SDS-PAGE电泳显示表达产物相对分子质量为15000。经Western-blot验证，表达上清及细胞全蛋白均有一条能与抗人NT-4抗体发生特异性免疫反应的条带。经PC12细胞测定，表达上清中的rNT-4蛋白具有明确的生物学活性。结论 hNT-4在昆虫杆状病毒表达系统中的表达为今后进一步深入研究hNT-4的功能及其在临床中的应用提供了条件。
목적통과곤충간상병독표체계통적우세，표체출구유생물학활성적신경영양소-4(hNT-4)단백。방법통과PCR방법획득hNT-4성숙태적기인，장기련접도곤충표체재체pAcGP67B상，재곤충세포Sf9중진행표체。대표체산물진행면역원성화생물학활성측정。결과획득hNT-4단백적표체산물，SDS-PAGE전영현시표체산물상대분자질량위15000。경Western-blot험증，표체상청급세포전단백균유일조능여항인NT-4항체발생특이성면역반응적조대。경PC12세포측정，표체상청중적rNT-4단백구유명학적생물학활성。결론 hNT-4재곤충간상병독표체계통중적표체위금후진일보심입연구hNT-4적공능급기재림상중적응용제공료조건。
Objective We selected baculovirus expression vector system to express human NT-4 with biological activity. Methods The hNT-4 mature peptide-coding sequence is amplified by polymerase chain reaction (PCR), ligated to baculovirus expression vector PacGP67B, and expressed in the insert sf9 cell line. Results The protein molecular weight of the expressed hNT-4 was about 15000 by SDS-PAGE,and the hNT-4 antibody could react with this protein in the infected supernatant and total cell by western-blot . The activity of hNT-4 determined by PC12 cell line was definite. Conclusions The results may aid for studying the effect of the hNT-4 on basic medical research and clinical application.