Abstract
BackgroundIdentification of genetic variants, including SNPs (Single Nucleotide Polymorphisms), CNVs (Copy Number Variations) and alternative splicing, within functional genes has received increasing attention in animal science research. HGF (Hepatocyte Growth Factor) is a very important growth factor that works as a mitogen or a morphogen during tissue growth, development and regeneration. However, to date, the functions of genetic variants within the bovine HGF gene, particularly their effects on mRNA expression, have not been determined well.ResultsThe present study aimed to perform association analysis between genetic variants and mRNA expression for the bovine HGF gene in Qinchuan cattle using various strategies, including PCR-RFLP (Restriction Fragment Length Polymorphism), qPCR (Quantitative Real-time quantitative PCR), TA cloning, DNA sequencing and bioinformatics analysis. A total of five SNPs were identified and only SV1 locus significantly affected HGF mRNA expression in fetal skeletal muscle (P < 0.05). Heterozygous genotype individuals showed significantly higher HGF expression (P < 0.05), which was significantly greater in the “CTCCAGGGTT” combined genotype than that in the “CCCCGGGGTT” combined genotype (P < 0.05). In addition, two alternative splicing variations, HGF-W and HGF-M, were identified, which resulted from alternative 3′ splice sites of exon 5, and HGF-W showed higher mRNA levels than HGF-M in all tissues.ConclusionIn summary, genetic variations within the HGF gene affected mRNA expression. These findings provide new insight into the molecular characteristics and functions of bovine HGF.
Highlights
Identification of genetic variants, including SNPs (Single Nucleotide Polymorphisms), Copy number variations (CNVs) (Copy Number Variations) and alternative splicing, within functional genes has received increasing attention in animal science research
Sample collection and genomic DNA/total RNA isolation Skeletal muscle samples from 48 male and 37 female fetal Qinchuan cattle were obtained for genomic DNA and total RNA isolation to detect the correlation between mutations and relative mRNA expression levels
The differences in expression levels between two alternative splicings in every tissue was Hepatocyte Growth Factor (HGF) mRNA expression based on genotypes in skeletal muscle of fetal bovine Firstly, the expression of HGF gene in female cattle was compared with that in male cattle using the t test, but no significant difference (P > 0.05) was observed
Summary
Identification of genetic variants, including SNPs (Single Nucleotide Polymorphisms), CNVs (Copy Number Variations) and alternative splicing, within functional genes has received increasing attention in animal science research. HGF (Hepatocyte Growth Factor) is a very important growth factor that works as a mitogen or a morphogen during tissue growth, development and regeneration. To date, the functions of genetic variants within the bovine HGF gene, their effects on mRNA expression, have not been determined well. Exonic SNPs have a direct effects on the properties of proteins, while SNPs within introns and untranslated region can affect the expression and splicing of mRNA [9, 10]. Numerous studies have shown that SNPs in the HGF gene are associated with normal growth and development [11,12,13]. It remains unclear that how these mutations affect gene expression, including the bovine HGF gene examined in this study.
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