CAJ | 학술논문

为选择湖羊小肠黏膜合适实时定量PCR的内参基因,本试验选用6只湖羊公羔,分别在42和84日龄各屠宰3只,研究候选内参基因甘油醛-3-磷酸脱氢酶( GAPDH)、磷酸甘油酸激酶基因1( PGK1)、18S rRNA和β-肌动蛋白( ACTB)在十二指肠、空肠、回肠黏膜发育过程中表达是否稳定. 结果表明:4 个候选基因都获得特异性扩增片段;在不同的小肠肠段黏膜中, PGK1和ACTB表达最稳定,在不同日龄4个候选基因表达都较稳定;ΔCt法和geNorm软件分析表明,ACTB表达最稳定. 综上,利用实时定量PCR研究湖羊小肠黏膜的内参基因最佳选择为ACTB.
위선택호양소장점막합괄실시정량PCR적내삼기인,본시험선용6지호양공고,분별재42화84일령각도재3지,연구후선내삼기인감유철-3-린산탈경매( GAPDH)、린산감유산격매기인1( PGK1)、18S rRNA화β-기동단백( ACTB)재십이지장、공장、회장점막발육과정중표체시부은정. 결과표명:4 개후선기인도획득특이성확증편단;재불동적소장장단점막중, PGK1화ACTB표체최은정,재불동일령4개후선기인표체도교은정;ΔCt법화geNorm연건분석표명,ACTB표체최은정. 종상,이용실시정량PCR연구호양소장점막적내삼기인최가선택위ACTB.
Six Hu lambs were used and three of them were slaughtered at 42 and 84 days of age separately to select a suitable reference gene for real time-PCR ( RT-PCR) of lamb intestinal mucosa. Stabilities of four can-didate housekeeping genes glyceraldehyde phosphate dehydrogenase ( GAPDH ) , phosphoglycerate kinase 1 ( PGK1) , 18S rRNA and β-actin ( ACTB) were measured in mucosa of duodenum, jejunum and ileum. The results showed as follow: four candidate housekeeping genes' specific gene fragments were detected; the ex-pression of PGK1 and ACTB turned out to be the most stable between mucosa of three small intestinal compart-ments; and all four candidate genes showed a constant expression between 42 and 84 days of age; the expres-sion of ACTB is the most stable gene according the analysis ofΔCt method and geNorm software. It is conclu-ded that ACTB is the best reference gene of RT-PCR for Hu lambs small intestinal mucosa.