CAJ | 학술논문

家蚕(Bombyx mori)的Nvd-Bm蛋白是一种与胆固醇7，8–脱氢酶相关的蛋白．根据NCBI中报道的家蚕中的胆固醇7，8–脱氢酶基因序列(GenBank 登录号：AB232986.1)，采用密码子优化及基因克隆技术，扩增获得胆固醇7，8–脱氢酶目标基因nvd-Bm，并构建了重组穿梭载体pPIC9K-nvd-Bm，通过电转化，成功构建了含有pPIC9K-nvd-Bm的毕赤酵母(Pichia pastoris)GS115异源真核表达工程菌株．经 SDS-PAGE 蛋白电泳分析表明，含有 pPIC9K-nvd-Bm 的毕赤酵母GS115表达菌株能够成功表达目标蛋白，为胆固醇的7，8–脱氢产物即7–脱氢胆固醇的生物转化奠定了基础．
가잠(Bombyx mori)적Nvd-Bm단백시일충여담고순7，8–탈경매상관적단백．근거NCBI중보도적가잠중적담고순7，8–탈경매기인서렬(GenBank 등록호：AB232986.1)，채용밀마자우화급기인극륭기술，확증획득담고순7，8–탈경매목표기인nvd-Bm，병구건료중조천사재체pPIC9K-nvd-Bm，통과전전화，성공구건료함유pPIC9K-nvd-Bm적필적효모(Pichia pastoris)GS115이원진핵표체공정균주．경 SDS-PAGE 단백전영분석표명，함유 pPIC9K-nvd-Bm 적필적효모GS115표체균주능구성공표체목표단백，위담고순적7，8–탈경산물즉7–탈경담고순적생물전화전정료기출．
Bombyx mori Nvd-Bm protein is associated with cholesterol 7,8-dehydrogenase. Based on DNA sequence en-coded cholesterol 7,8-dehydrogenase reported on the NCBI(GenBank:AB232986.1),cholesterol 7,8-dehydrogenase gene was cloned from Bombyx mori. Through codon optimization,cholesterol 7,8-dehydrogenase gene was obtained and the op-timized gene was cloned into a yeast expression vector. Heterologously expressed target gene in P. pastoris GS115 resulted in the creation of engineered P. pastoris GS115/pPIC9K-nvd-Bm strains. Expression of the target protein in the engineered P. pastoris GS115/pPIC9K-nvd-Bm strains was demonstrated by SDS-PAGE. This study provided a foundation for the biologi-cal synthesis of 7-dehydrocholesterol.