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1.
Anim Biotechnol ; 31(1): 81-85, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-30570382

RESUMO

Glycosylphosphatidylinositol-anchored high-density lipoprotein-binding protein 1 (GPIHBP1) is among the many candidate genes for regulating milk production traits in dairy cattle that have been identified via quantitative trait locus (QTL) mapping and genome-wide association studies (GWAS). Our previous studies confirmed that a G-to-A mutation at chr14:2553998 is the main cause of GPIHBP1-related effects on milk fat content. In this study, we discovered that GPIHBP1 may be a strong candidate gene for the regulation of milk protein traits. We performed overexpression and RNAi experiments to assess GPIHBP1 in bovine primary mammary epithelial cells (BMECs) and identified mRNA expression patterns of several important milk protein-related genes using real-time quantitative PCR. After the transient transfection of BMECs with GPIHBP1, the transcription levels of casein genes (CSN1S1, CSN1S2, CSN2, and CSN3) and lactoferrin (LTF) decreased, whereas beta-lactoglobulin (LGB) expression increased. The GPIHBP1 RNAi experiment produced changes in gene expression that were completely opposite to those observed in the GPIHBP1 overexpression experiment. Furthermore, among the assessed genes, CSN3, LTF, and LGB exhibited significant changes in mRNA expression (p < 0.05). The findings of this study show that bovine GPIHBP1 is involved in the process of milk protein biosynthesis and may be considered as a functional gene for the milk protein yield trait.


Assuntos
Bovinos/genética , Regulação da Expressão Gênica , Proteínas do Leite/genética , Leite/química , Receptores de Lipoproteínas/metabolismo , Animais , Caseínas/metabolismo , Bovinos/metabolismo , Contagem de Células/veterinária , Mapeamento Cromossômico/veterinária , Indústria de Laticínios , Células Epiteliais/metabolismo , Feminino , Glândulas Mamárias Animais/metabolismo , Mutação , Fenótipo , Receptores de Lipoproteínas/genética
2.
Asian-Australas J Anim Sci ; 33(11): 1725-1731, 2020 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-32106650

RESUMO

OBJECTIVE: An initial RNA-Sequencing study revealed that UDP-galactose-4-epimerase (GALE) was one of the most promising candidates for milk protein concentration in Chinese Holstein cattle. This enzyme catalyzes the interconversion of UDP-galactose and UDP-glucose, an important step in galactose catabolism. To further validate the genetic effect of GALE on milk protein traits, genetic variations were identified, and genotypes-phenotypes associations were performed. METHODS: The entire coding region and the 5'-regulatory region (5'-UTR) of GALE were re-sequenced using pooled DNA of 17 unrelated sires. Association studies for five milk production traits were performed using a mixed linear animal model with a population encompassing 1,027 Chinese Holstein cows. RESULTS: A total of three variants in GALE were identified, including two novel variants (g.2114 A>G and g.2037 G>A) in the 5'-UTR and one previously reported variant (g.3836 G>C) in an intron. All three single nucleotide polymorphisms (SNPs) were associated with milk yield (p<0.0001), fat yield (p = 0.0006 to <0.0001), protein yield (p = 0.0232 to <0.0001) and protein percentage (p<0.0001), while no significant associations were detected between the SNPs and fat percentage. A strong linkage disequilibrium (D' = 0.96 to 1.00) was observed among all three SNPs, and a 5 Kb haplotype block involving three main haplotypes with GAG, AGC, and AGG was formed. The results of haplotype association analyses were consistent with the results of single locus association analysis (p<0.0001). The phenotypic variance ratio above 3.00% was observed for milk protein yield that was explained by SNP-g.3836G >C. CONCLUSION: Overall, our findings provided new insights into the polymorphic variations in bovine GALE gene and their associations with milk protein concentration. The data indicate their potential uses for marker-assisted breeding or genetic selection schemes.

3.
Genes (Basel) ; 10(2)2019 01 30.
Artigo em Inglês | MEDLINE | ID: mdl-30704154

RESUMO

Heat shock 70 kDa protein 8 (HSPA8) and erb-b2 receptor tyrosine kinase 2 (ERBB2) were the promising candidates for milk protein concentration in dairy cattle revealed through previous RNA sequencing (RNA-Seq) study. The objective of this post-RNA-Seq study was to confirm genetic effects of HSPA8 and ERBB2 on milk protein concentration in a large Chinese Holstein population and to evaluate the genetic effects of both genes on other milk production traits. There were 2 singlenucleotide polymorphisms (SNPs) identified for HSPA8 and 11 SNPs for ERBB2 by sequencing 17 unrelated Chinese Holstein sires. The SNP-rs136632043 in HSPA8 had significant associations with all five milk production traits (p = 0.0086 to p < 0.0001), whereas SNP-rs132976221 was remarkably associated with three yield traits (p < 0.0001). Nine (ss1996900615, rs109017161, rs109122971, ss1996900614, rs110133654, rs109941438, rs110552983, rs133031530, and rs109763505) of 11 SNPs in ERBB2 were significantly associated with milk protein percentage (p = 0.0177 to p < 0.0001). A 12 Kb haplotype block was formed in ERBB2 and haplotype associations revealed similar effects on milk protein traits. Our findings confirmed the significant genetic effects of HSPA8 and ERBB2 on milk protein concentration and other milk production traits and SNP phenotypic variances above 1% may serve as genetic markers in dairy cattle breeding programs.


Assuntos
Bovinos/genética , Proteínas de Choque Térmico HSC70/genética , Proteínas do Leite/genética , Polimorfismo de Nucleotídeo Único , Receptor ErbB-2/genética , Animais , Haplótipos , Proteínas do Leite/metabolismo , Característica Quantitativa Herdável
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