Linkage disequilibrium and functional analysis of PRE1 insertion together with SNPs in the promoter region of IGFBP7 gene in different pig breeds
Polymorphisms in regions upstream of transcription initiation site may modify the transcriptional activity of target genes by changing promoter activity. This study aims to determine whether or not polymorphisms at porcine IGFBP7 promoter region affect gene expression. In this study, eight SNPs and one PRE1 insertion in this region were first confirmed. The PRE1 insertion was widespread in 20 Chinese indigenous breeds, but was not observed in three commercial breeds. A perfect linkage disequilibrium, consisting of six of those SNPs and a PRE1, was observed with two haplotypes (h1 and h2) in five pig breeds. The h1 haplotype had an overwhelming superiority distribution in Large White, Landrace, and Bama mini-pig; in turn, the h2 only existed in the PRE1 presence breeds. As the haplotypes and PRE1 were located at gene promoter regions, we further investigated the transfection of plasmids with three different fragments of IGFBP-7 promoter region (H1, H2, RF). The CMV promoter of the pEGFP-N1 was substituted by these three different fragments, respectively. Different transcriptional and translational activities of EGFP in PK-15 cells were observed in these three recombinant plasmids by quantitative real-time PCR and flow cytometric analysis. The results indicated that H1 had the higher transcriptional and translational activities of EGFP as compared to the H2 (P < 0.05, P < 0.05). As compared to the RF group, EGFP mRNA expression level was significantly higher in H1 groups (P < 0.05). The IGFBP-7 promoter polymorphisms detected in this study may be important functional variants and potential genetic markers for pig population genetic study.
KeywordsPRE1 SNP LD Haplotype Promoter activity
This work was supported by the National Natural Science Foundations of China (31672514 and 31772699) and Science and Technology Development Plan of Jilin Province (20170101024JC).
Compliance with ethical standards
Conflict of interest
The authors declare that they have no competing interests.
- Fontanesi L, Scotti E, Buttazzoni L, Dall’Olio S, Russo V (2014) Investigation of a short interspersed nuclear element polymorphic site in the porcine Vertnin gene: allele frequencies and association study with meat quality, carcass and production traits in Italian large white pigs. Ital J Anim Sci 13(1):3090CrossRefGoogle Scholar
- Li W et al (2005) Isolation of differentially expressed genes in double-muscling large white pig by SSH and Q-PCR strategy. Prog Biochem Biophys 32:353–358Google Scholar