不同定量方法检测猪背最长肌中ssc-miR-27a表达研究.docVIP

  不同定量方法检测猪背最长肌中 ssc-miR-27a 表达研究# 虞德兵* 5 10 （南京农业大学动物科技学院，南京 210095） 摘要：miRNA 是一类长度为 21-25nt 的非编码 RNA，通过与靶基因的 3′UTR 区结合来 调控基因表达，在调节生物体生长、发育、疾病发生过程等方面发挥重要作用。本研究采 用 step-loop PCR 法和 Poly(T) adaptor 法，结合 real time PCR 技术检测猪 miR-27a 在二花脸 猪和长大杂交猪高、低肌内脂肪组个体背最长肌中的表达量, 并进行表达量的相关性分 析，以验证两种方法的可靠性。结果表明，两种方法获得两组基因表达量值呈高度相关 （R2=0.9349）,且呈显著性相关(p 0.05)。miR-27a 在同一品种高低肌内脂肪的猪骨骼肌内 有表达差异(p0.01)，且在不同品种猪间也存在表达差异(p 0.05)。 关键词：猪；miR-27a；背最长肌 中图分类号：S852 15 Study on the expression of miR-27a by real time PCR in Pig Longessi Dorsi YU Debing (College of Animal Science and Technology, Nanjing Agricultural University, NanJing 210095) 20 25 30 Abstract: MicroRNAs (miRNAs) were a recently discovered class of endogenous, small non-encoding RNAs with the length of 21-25 nucleotides that play important roles in the negative regulation of gene expression by base pairing to complementary sites in the target messenger RNA (mRNA) 3’-untranslated region (UTR),it has been confirmed that they regulate complicated biological behaviors, such as cell differentiation, proliferation and death. The step-loop PCR and poly(T) adaptor,and Combined with real-time PCR method were used to detect relative quantity of miR-27a in Longissimus Dorsi of Erhua lian and Large White×Landrace pigs,at the same time, the relationship analysis of the expresion from different methods were examined, so as to proclaim the reliability of two methods. The results showed that the expression level of miR-27a in low IMF group was more that it in high IMF group from the Large White×Landrace pigs (P 0.01), otherwise, the expression level of MSTN mRNA in low IMF group was more that it in high IMF group from the Erhua lian pigs (P 0.01)，in contrast to White×Landrace pigs(P 0.05). All the results suggested that there was a real relationship of gene expression between step-loop PCR and poly(T) adaptor(P 0.05). Keywords: pig; miR-27a; Longissimus Dorsi 35 0 引言 MicroRNA(miRNA) 是一类长度约 22nt 的单链非编码小分子 RNA, 在生物的进化过程中 高度保守，通过与靶 mRNA3 端非翻译区(