CAJ | 학술논문

以水稻α-半乳糖苷酶重组质粒pET32a+-84411为模板，通过PCR反应条件的筛选，琼脂糖凝胶电泳检测，扩增出条带清晰、单一的水稻α-半乳糖苷酶（BAC84411.1）目的基因，通过双限制性内切酶的酶切插入到酵母表面展示载体pYD1中，此目的基因与载体以摩尔数比为4∶1的比例进行连接。并以将重组质粒转化到大肠杆菌JM109中，并提取到了阳性重组子质粒，以备转化到酵母表达菌株EBY100中，诱导其表达α-半乳糖苷酶。
이수도α-반유당감매중조질립pET32a+-84411위모판，통과PCR반응조건적사선，경지당응효전영검측，확증출조대청석、단일적수도α-반유당감매（BAC84411.1）목적기인，통과쌍한제성내절매적매절삽입도효모표면전시재체pYD1중，차목적기인여재체이마이수비위4∶1적비례진행련접。병이장중조질립전화도대장간균JM109중，병제취도료양성중조자질립，이비전화도효모표체균주EBY100중，유도기표체α-반유당감매。
Riceα-galactosidase (BAC84411) recombinant plasmid pET32a+-84411 as template, PCR amplifica-tion conditions and the reaction system were screened. The amplified fragment were detected by agarose gel electrophoresis, a clear and single band of the target gene was obtained. And the gene was inserted into the yeast surface display vector pYD1 by double restriction endonuclease. the insert and the vector ligased by moles ratio of 4∶1. Then the ligation mixture was transformed into E.coli JM109, and the positive recombinant plasmid was identified. The obtained pYD1-84411plasmid would be used for inducing α-Galactosidase expression in yeast strains EBY100.