CAJ | 학술논문

神经肽Y (Neuropeptide Y, NPY)被普遍认为是一种重要的促食因子,在调节鱼类的摄食行为方面起着至关重要的作用。为进一步研究牙鲆 NPY 蛋白的生物学功能,作者克隆了牙鲆 NPY 成熟肽序列(m-NPY)及含有信号肽的全长序列(f-NPY),利用原核表达系统分别进行体外重组表达,筛选出诱导剂IPTG最佳诱导浓度为0.8 mmol/L及最佳诱导时间3 h；此外,根据牙鲆NPY第49-64位氨基酸序列制备了多克隆抗体并通过Western blot验证该多抗能够有效检验重组 NPY及牙鲆体内NPY的表达。研究结果为研究重组牙鲆NPY蛋白在牙鲆水产养殖产业中的应用及检测提供了依据。
신경태Y (Neuropeptide Y, NPY)피보편인위시일충중요적촉식인자,재조절어류적섭식행위방면기착지관중요적작용。위진일보연구아평 NPY 단백적생물학공능,작자극륭료아평 NPY 성숙태서렬(m-NPY)급함유신호태적전장서렬(f-NPY),이용원핵표체계통분별진행체외중조표체,사선출유도제IPTG최가유도농도위0.8 mmol/L급최가유도시간3 h；차외,근거아평NPY제49-64위안기산서렬제비료다극륭항체병통과Western blot험증해다항능구유효검험중조 NPY급아평체내NPY적표체。연구결과위연구중조아평NPY단백재아평수산양식산업중적응용급검측제공료의거。
Neuropeptide Y (NPY) plays a key role in regulation of food intake in fish, which is essential for aqua-culture. In order to investigate the effects of NPY on olive flounder (Paralichthys olivaceus), two types of flounder NPY gene, full length NPY (f-NPY) with signal peptide and mature peptide NPY (m-NPY) without the signal, were cloned into expression vector pPROEXTM HTa and expressed in E. coli BL21 (DE3) respectively. The recombinant NPY proteins were induced by IPTG. The optimal concentration of IPTG was 0.8 mmol/L and the optimal induction time was 3 h. In addition, the NPY polyclonal antibody was prepared according to its amino acid sequence. Western blot analysis confirmed that both the recombinant NPY and the flounder endogenous NPY protein could be detected by the polyclonal antibody. In summary, our study will provide basis for further study on the application of recom-binant olive flounder NPY in aquaculture.