A SYBR Green I-based qRT-PCR Assay for Detecting Duck BMPs

  Objective   An assay for detecting mRNA transcription of bone morphogenetic proteins (BMPs) in ducks using the SYBR Green Ⅰ-based qRT-PCR was developed.

  Methods   Specific primers were designed and synthesized according to the nucleotide sequences of duck BMPs in GenBank. The PCR amplified products were cloned into pMD-18-T vector, and the recombinant plasmids DNA used to establish standard curves. Specificity, sensitivity, and repeatability of the methodology were examined.

  Result   The newly developed qRT-PCR assay showed singe specific melting peaks of BMP1, BMP2, BMP3, BMP4, BMP5, BMP6, BMP7, BMP8a, BMP10, and BMP15 separately with correlation coefficients (R²) higher than 0.998. The coefficients of variation within and between groups were less than 1%. The mRNA expressions of the BMPs were detected in different tissues of hybrid Muscovy ducks. The expressions of BMP1, BMP2, BMP5, and BMP7 in the heart were significantly higher than those in other tissues at 5.5×10⁴, 1.1×10⁵, 5.1×10⁵, and 7.7×10⁵ copies·μL⁻¹, respectively. BMP6 and BMP10 were highly expressed in the liver at 7.5×10⁴ and 7.6×10³ copies·μL⁻¹, respectively, while BMP3 in Bursa of Fabricius, BMP4 in the thymus, BMP8a in the beak, and BMP15 in the spleen at 1.5×10⁵, 2.1×10⁴, 1.8×10³, and 3.3×10³ copies·μL⁻¹, respectively.

  Conclusion  The newly developed qRT-PCR assay for the determination of mRNA expression of BMPs in ducks was specific, sensitive, repeatable, and applicable.