CAJ | 학술논문

三磷酸甘油醛脱氢酶(GAPDH)是一种糖酵解酶,在脊椎动物的所有组织均有稳定的表达,经常作为看家基因来研究基因的表达,本研究分析了鸭GAPDH基因序列,设计了一对引物,利用PCR,RT-PCR技术扩增得到了浙东白鹅的GAPDH基因的DNA序列为235 bp,mRNA序列为209 bp,作为参比基因应用于荧光定量PCR来研究目标基因的表达,目标基因在mRNA水平上的表达量可通过参比基因校正cDNA加入量的不同来研究,荧光定量PCR显示了以cDNA为模板,此引物扩增时Ct具有单一的溶解曲线,说明其表达稳定,可以作为参比基因用于浙东白鹅基因的定量表达研究.
삼린산감유철탈경매(GAPDH)시일충당효해매,재척추동물적소유조직균유은정적표체,경상작위간가기인래연구기인적표체,본연구분석료압GAPDH기인서렬,설계료일대인물,이용PCR,RT-PCR기술확증득도료절동백아적GAPDH기인적DNA서렬위235 bp,mRNA서렬위209 bp,작위삼비기인응용우형광정량PCR래연구목표기인적표체,목표기인재mRNA수평상적표체량가통과삼비기인교정cDNA가입량적불동래연구,형광정량PCR현시료이cDNA위모판,차인물확증시Ct구유단일적용해곡선,설명기표체은정,가이작위삼비기인용우절동백아기인적정량표체연구.
Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is a glycoytic enzyme, which is active in all vertebrate tissues and frequently used as a housekeeping gene in expression studies. In this study, the mRNA sequence of glyceraldehyde-3-phosphate dehydrogenase in duck was analyzed by designing a pair of primers and ampified in eastern Zhejiang white geese (a native Chinese geese breed) as a control gene in real time PCR. The mRNA levels of the target gene were measured relatively to their respective templates and were normalized to reference genes,to control the amount of cDNA loaded into the reaction. The sequences of genomic DNA and cDNA of goose glyceraldehydes-3-phophate dehydrogenase were 235 bp and 209 bp, respectively. Real time PCR showed a specific Ct using cDNA as template, and the primers showed a single melt peak. So the gene expression study was reliable when glyceraldehyde-3-phosphate dehydrogenase was used as an internal reference gene.