Analysis of Promoter Activity of Porcine SIM1 Gene and Its Expression Pattern

doi:10.7668/hbnxb.2015.02.002

Abstract

Abstract: To investigate the transcriptional regulation mechanism and expression pattern of porcine Single-minded 1 (SIM1) gene.5' promoter fragment was amplified with PCR method and the potential transcription factor binding sites were predicted with bioinformatics method;Eight promoter segments with different length were obtained by promoter deletion analysis and cloned into pGL3-Enhancer vector, then the promoter activities were determined in transiently transfected 293T, MGC803 and Bel7402 cells by the dual-luciferase assay system, respectively.Meanwhile, SIM1 protein expression pattern was detected in seven tissues using Western Blot (WB) method.The results showed that all these SIM1 promoters presented activities in 293T, MGC803 and Bel7402 cells, and the lowest in 293T cells.The region of -699 — -489 bp contained the key cis-regulatory elements and further bioinformatics analysis found some key transcription factor binding sits, such as Smad, CEBPα and PAX6.WB experiments revealed that, besides its known expression in central nervous system, SIM1 protein could be detected in all examined tissues.Grayscale analysis showed a strong SIM1 protein expression in brain, hypothalamus and testes, a weak to moderate expression in muscle, thyroid gland, subcutaneous fat and liver.These results indicated that SIM1 gene may play important role in nerve cell development, fatness metabolism and gonads development.

Key words: Pig, SIM1 gene, Promoter activity, Expression pattern

CLC Number:

CHEN Zhe, LEI Ming-ming, SHI Zhen-dan. Analysis of Promoter Activity of Porcine SIM1 Gene and Its Expression Pattern[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2015, 30(2): 6-11. doi: 10.7668/hbnxb.2015.02.002.

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