Selection of Appropriate Reference Genes for RT-qPCR Analysis in Different Tissue with Different Development Periods in Dapulian Pigs

doi:10.7668/hbnxb.2018.S1.008

Abstract

Abstract: To obtain the accurate RT-qPCR results,the expression levels of genes of interest should be normalized with appropriate and optimal numbers of reference genes. In this study,using two excel-based programs,geNorm and NormFind,we assessed the expression stability of eight well-known candidate reference genes(ACTB,B2M,GAPDH,RPL4,SDHA,TBP,YWHAZ and PPIA) in eight body tissues(heart,liver,spleen,lung,kidney,intestines,muscle and peripheral blood) with different development periods of Dapulian pigs. Combination analysis of two programs showed that the stable and appropriate reference genes were TBP, PPIA and YWHAZ in heart; TBP and PPIA in liver,kidney,intestines and muscle; TBP and RPL4 in spleen; TBP, RPL4, PPIA and ACTB in lung; ACTB, YWHAZ and GAPDH in peripheral blood. The difference of suitable number and composition reference genes in eight body tissues of Dapulian indicated that the selection studies of optimal reference genes were essential in RT-qPCR analysis. The results provided reliable reference genes for expression studying of various genes related to diseases,meat quality,litter size and reproduction in Dapulian pigs.

Key words: Dapulian pigs, RT-qPCR, Reference genes, geNorm, NormFind

CLC Number:

Q78
S828

WANG Yanping, ZHU Rongsheng, WANG Huaizhong, WANG Cheng, HU Hongmei. Selection of Appropriate Reference Genes for RT-qPCR Analysis in Different Tissue with Different Development Periods in Dapulian Pigs[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2018, 33(S1): 45-53. doi: 10.7668/hbnxb.2018.S1.008.

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