关键词: 猪, PFK-2/FBPase-2 mRNA, SYBR Green I, 荧光定量实时-聚合酶链式反应

Abstract:

In order to explore the relationship between PFK-2/FBPase-2 mRNA transcription level and postmortem meat
quality, a pair of primers was designed and β-actin was chosen as housekeeper gene in this study according to swine PFK-
2/FBPase-2 mRNA sequences available in GenBank (NM_001143721.1). Under the optimal experimental conditions, a
real-time fluorescent quantitative reverse transcription-polymerase chain reaction (RT-PCR) assay was established to detect
swine PFK-2/FBPase-2 gene. The results showed that both the correlation coefficient (R2) of the standard curves of PFK-
2/FBPase-2 and the housekeeper gene β-actin were more than 0.999, the sensitivity was high and detection limits of the
two genes were 10 copies/μL. In addition, the specificity was high and the amplification products were specifically single
peaks. Meanwhile the amplification efficiency of PFK-2/FBPase-2 (104.5%) was consistent with that of β-actin (104.0%).
According to the extent of PSE meat production, the longissimus dorsi muscles of Taihu, Duroc and Duroc× (Landrace ×
Yorkshire) were selected to measure the transcription level of PFK-2/FBPase-2 mRNA at 45 min in different breeds by the
established method. The results showed that Taihu had the highest PFK-2/FBPase-2 mRNA transcript level, followed by Duroc, the
lowest was Duroc × (Landrace × Yorkshire), and there was significant difference among these three breeds (P ＜ 0.05).

Key words: swine, PFK-2/FBPase-2 mRNA, SYBR Green I, real time-polymerase chain reaction (RT-PCR)