A SYBR Green I-based qRT-PCR Assay for Detecting Duck <i>BMP</i>s

LIU Hongwei; LIN Chang; WANG Shao; XIAO Shifeng; CHENG Xiaoxia; ZHENG Min; CHEN Shaoying; ZENG Xiancheng; CHEN Shilong

doi:10.19303/j.issn.1008-0384.2022.012.001

Volume 37 Issue 12

Dec. 2022

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Article Navigation > Fujian Journal of Agricultural Sciences > 2022 > 37(12): 1509-1518

LIU H W, LIN C, WANG S, et al. A SYBR Green I-based qRT-PCR Assay for Detecting Duck BMPs [J]. Fujian Journal of Agricultural Sciences，2022，37（12）：1509−1518 doi: 10.19303/j.issn.1008-0384.2022.012.001

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LIU H W, LIN C, WANG S, et al. A SYBR Green I-based qRT-PCR Assay for Detecting Duck BMPs [J]. Fujian Journal of Agricultural Sciences，2022，37（12）：1509−1518 doi: 10.19303/j.issn.1008-0384.2022.012.001

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

doi: 10.19303/j.issn.1008-0384.2022.012.001

LIU Hongwei^{1, 2
,},
LIN Chang^{1, 2},
WANG Shao^{2, 3},
XIAO Shifeng^{2, 3},
CHENG Xiaoxia^{2, 3},
ZHENG Min^{2, 3},
CHEN Shaoying^{2, 3},
ZENG Xiancheng^{1
,
,},
CHEN Shilong^{2, 3
,
,}

1.
College of Animal Sciences (College of Bee Science), Fujian Agricultural and Forestry University, Fuzhou, Fujian　350002, China
2.
Institute of Animal Husbandry and Veterinary Medicine, Fujian Academy of Agriculture Sciences, Fuzhou, Fujian　350013, China
3.
Putian Branch, Fujian Academy of Agricultural Sciences, Fuzhou, Fujian　351100, China

Received Date: 2022-09-28
Rev Recd Date: 2022-12-01

Available Online: 2022-12-28

Publish Date: 2022-03-28

Abstract

Abstract

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.
- Duck,
- bone morphogenetic proteins,
- qRT-PCR,
- SYBR Green I

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References(23)

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