合浦珠母贝实时定量PCR内参基因的稳定性比较

Stability comparison of reference genes of Pinctada fucata by real-time qPCR

  • 摘要: 以合浦珠母贝(Pinctada fucata)为研究对象, 应用实时荧光定量PCR技术检测了甘油醛-3-磷酸脱氢酶(GAPDH)、β-肌动蛋白(β-actin)和18S核糖体rRNA(18S rRNA)3个管家基因在合浦珠母贝不同组织、性腺发育时期、胚胎发育时期mRNA水平的表达情况。同时比较了BestKeeper、geNorm和NormFinde软件对试验数据处理的差异, 从中选出合适的内参基因。结果表明β-actin在不同组织和胚胎发育不同阶段表达最稳定, 18S rRNA在性腺发育不同时期表达最稳定。

     

    Abstract: We analyze the expression stability of the housekeeping genes in Pinctada fucata, including Glyceraldehyde-3-Phosphate Dehydrogenase (GAPDH), beta-actin (β-actin) and 18S rRNA (18S), by quantitative real-time polymerase chain reaction (qRT-PCR) in different tissues, at different stages of gonadal development and embryonic development.Besides, we select the suitable reference genes by using softwares of BestKeeper, geNorm and NormFinder algorithms.The results indicate that β-actin is the most stable reference gene for different tissues and different embryonic developmental stages, while 18S is the most stable reference gene for different gonadal developmental stages.

     

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