A Novel Missense Single Nucleotide Polymorphism in the GREM1 Gene is Highly Associated with Higher Reproductive Traits i
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A Novel Missense Single Nucleotide Polymorphism in the GREM1 Gene is Highly Associated with Higher Reproductive Traits in Awassi Sheep Faris S. Imran, et al. [full author details at the end of the article] Received: 7 April 2020 / Accepted: 29 September 2020 © Springer Science+Business Media, LLC, part of Springer Nature 2020
Abstract GREM1 (gremlin1) is a known inhibitor for BMP15 (bone morphogenetic protein 15) family, but its genetic diversity in sheep is unknown. The present study was conducted to analyze the polymorphism of GREM1 gene using PCR- single-strand conformation polymorphism (SSCP) and DNA sequencing methods and to assess the possible association of GREM1 gene polymorphism with reproductive traits in Awassi ewes. A total of 224 ewes, 124 producing singles and 100 producing twins, were included in the study. Two SSCP patterns were detected in two amplified loci within the exon 2. Two exonic novel single nucleotide polymorphism (SNP)s were identified, c.74 T > G (the silent SNP p.Met123 =) and c.30 T > A with (the missense SNP p.Ile237Phe). Statistical analyses indicated a non-significant (P > 0.05) association of p.Met123 = with the analyzed reproductive traits of fecundity, prolificacy, litter size, and twinning rate. Meanwhile, p.Ile237Phe SNP exhibited a highly significant (P 3.5–5), and eijkl is the random error associated with Yijkl observation and assumed to be normally distributed with mean = 0 and variance = σ2e. Means were compared using the Tukey-Krammer test
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Biochemical Genetics
with a significance level of (P G, only in one of the SSCP variants. According to this nucleic acid substitution, the detected SSCP variants were assigned TT and TG genotypes, for the observed homozygous T/T and the heterogeneous T/G patterns that were ,respectively, detected at 77th position. This nucleic acid substitution was positioned in methionine residue at the 123rd residue within gremlin1. Expasy translate software indicated a silent effect of this SNP (p.Met123 =). Concerning 318 bp amplicons, sequencing reactions validated the PCR-SSCP heterogeneity by detecting additional novel SNP, c.30 T > G in one of the SSCP variants. A homozygous T/T status at 30th position in the two bands pattern and a heterogeneous T/A status at the same position in the three bands pattern were detected by the analyzed electropherograms (Fig. 1c). Accordingly, the detected SSCP variants were assigned TT and TA genotypes, respectively. This nucleic acid substitution was positioned in isoleucine residue at the 237th residue within gremlin1. Expasy translate software showed a missense effect of this SNP, in which an interesting amino acid substitution occurred from isoleucine to phenylalanine at the 237th position of the mature gremlin protein (p.Ile237Phe) (Fig. 1d). In addition to c.77 T > G and c.30 T > A, sequencing reactions identified two other nucleic acid variations: c.74 T > G (p.Ala122Pro) and c.280 T > C (p.Gln153 =) in the coding region of the GREM1 gene. However, both p.Ala122Pro and p.Gln153 = SNPs were det
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