Abstract
Selective breeding has strongly reduced the genetic diversity in livestock species and contemporary breeding practices exclude potentially beneficial rare genetic variation from the future gene pool. Here we test whether important traits arising by new mutations can be identified and rescued in highly selected populations. We screened milks from 2.5 million cows to identify an exceptional individual which produced milk with reduced saturated fat content and improved unsaturated and omega-3 fatty acid concentrations. The milk traits were transmitted dominantly to her offspring and genetic mapping and genome sequencing revealed a new mutation in a previously unknown splice enhancer of the DGAT1 gene. Homozygous carriers show features of human diarrheal disorders and may be useful for the development of therapeutic strategies. Our study demonstrates that high-throughput phenotypic screening can uncover rich genetic diversity even in inbred populations and introduces a novel strategy to develop novel milks with improved nutritional properties.
Similar content being viewed by others
Introduction
Livestock selection for the improved production of milk, meat, skin and fiber has influenced the evolution of human society and animal breeds1,2. Traditional selection based on ancestral performance is currently being supplanted by “genomic selection” based principally on the summation of genome-wide marker or haplotype associations with ancestral performance parameters3. While both industrial processes have provided considerable genetic gain, application of either methodology excludes rare alleles or new mutations with significant phenotypic impact from the future gene pool.
Haldane4 estimated the mutation rate for hemophilia in humans to be approximately 10−5 and subsequent studies have confirmed the rate of new mutations per gene to range from 10−6 to 10−4 per generation5,6,7. Due to the comparable sizes of human and cow genomes8,9, we hypothesized that if novel mutations were to arise in the bovine at a comparable rate, rare alleles for extreme phenotypes should be detectable provided that the sample size is above one million and an appropriate screening protocol for the rare trait is available. We also hypothesized that variations in these genetically and biochemically complex components could be proxies for individually refined milk compositions, including variations in individual milk fatty acids.
We have tested our hypothesis using a large and well-characterized bovine population. Our aim was to discover new alleles that are responsible for milks with extremely low or high milk fat or protein content.
Results
We screened the lactation records of 2.5 million cows and identified 29 individuals with milks deviating by more than 3.5 standard deviations (SD), over repeated milk tests and lactations, from the breed- and age-adjusted means for protein or fat content. Of these, Holstein-Friesian cow ‘363’ produced milk with average fat content of 2.81% ± 0.10% (mean ± SD), 3.9 SD below the breed mean of 4.52%. Milk protein concentration of cow 363 was 1 SD lower than the population group mean, while mean seasonal milk production of 6223 ± 925 L was 3.0 SD above the breed average. At selection, cow 363 was 8 years old and had completed five lactations, indicating normal fertility and survival rate. Information provided by her breeder and lifetime owner confirmed normal history and health status and the cow showed no abnormalities on inspection (Fig. 1b). The 74,618 paternal half-sisters of cow 363, farmed throughout New Zealand, produced milks with an average fat content of 4.4 ± 0.7%. Milks from cow 363's deceased dam and her maternal half-sister contained 3.98 ± 0.15% and 5.60 ± 0.3% fat, respectively.
Detailed milk composition analysis revealed a substantial improvement in the ratio of saturated : unsaturated milk fat. Saturated fat content was reduced by 3–4 SD, while mono- and polyunsaturated fatty acids were 2–3 SD above the New Zealand average10 (Supplementary Table 1). In contrast, content and composition of casein and whey proteins were within one 1 SD of the national breed average (Supplementary Table 2). The extreme milk fat content and milk fat composition phenotypes remained unchanged after cow 363 was relocated from her home farm to our research farm, indicating a negligible influence of environmental factors.
To assess the genetic basis of the extreme milk fat phenotypes, we generated four additional sons and seven daughters of cow 363 (Fig. 1a). Three daughters produced milks with an average fat content of 2.62% ± 0.09% and fatty acid compositions similar to cow 363; in contrast, average fat content (4.14% ± 0.37%) and fatty acid composition of milks from her other four daughters were similar to unrelated control cows (Fig. 1c, Supplementary Table 3). When unrelated Holstein-Friesian dams were bred with semen from the five sons of cow 363, 18 of the 50 daughters sired by three of the sons produced milks with fat content similar to that of founder cow 363. Milks from their paternal half-sisters and from the 42 daughters of the other two bull sons of cow 363, were similar to that of the breed average (Fig. 1a). Similar to the founder, no other phenotypes were apparent in her offspring. Taken together, these results suggested that a heterozygous genetic variation with a dominant effect was responsible for the milk fat phenotypes of cow 363.
Genotyping of 56,000 single-nucleotide markers in the pedigree of cow 363 and genome-wide association analysis identified a single locus associated at a genome-wide level of significance [p-value ≤ 1.27 × 10−6] (Fig. 2). This locus harbors the DGAT1 gene encoding diacylglycerol O-acyltransferase 1, which catalyzes the terminal reaction in triglyceride synthesis11,12,13. Sequencing of the DGAT1 locus of cow 363 revealed a heterozygous, non-synonymous A>C transversion in exon 16 (Fig. 3a). This variation was present in all females producing milks with less than 3% fat and in the three bulls producing offspring with reduced milk fat content but was absent in all other offspring of cow 363, her sire and her grand sires. Moreover, the variant was absent in 185 bulls that have sired the majority of the New Zealand dairy cows. As historical records showed that the milk fat phenotype of cow 363's deceased dam was typical of the breed, the mutation has likely occurred de novo. Cow 363 was homozygous for the common Ala 232 variant of DGAT114,15.
Quantitative PCR amplification polymerase and cDNA sequencing revealed that the mutation disrupts splicing and causes exon 16 skipping. In liver and mammary gland biopsies from heterozygous animals, transcripts lacking the 63-nucleotide exon 16 were as abundant as full-length transcripts (Fig. 3b). In contrast, only low levels of full-length transcripts were detectable in liver biopsies from homozygous mutants (Fig. 3b,c). Analysis of the wild-type sequence of exon 16 identified the putative consensus exonic splicing enhancer16,17 motif 5′-ATGATG overlapping the mutation (Fig. 3a).
To assess the effect of the mutation on enzyme activity, we expressed wild-type and mutant DGAT1 cDNAs in a bakers' yeast strain lacking endogenous diacylglycerol acyltransferase activity18. In contrast to the full-length variants, the mutant enzyme lacking the 21 amino acids encoded by exon 16 was unable to transfer oleic acid from CoA to diacylglycerol (Fig. 4).
These results show that a novel single nucleotide substitution in an exonic splicing enhancer of the DGAT1 gene induces exon 16 skipping and results in enzymatically inactive diacylglycerol O-acyltransferase 1. The mutation also reveals a link between triglyceride synthesis capacity and milk fat saturation and provides an important clue to the role of DGAT1 in milk fat saturation19.
Cows heterozygous for the new mutation thrived in a pastoral environment and displayed normal growth, survival and fertility rates. In contrast, homozygous calves exhibited scouring (non-bloody watery diarrhea), slow growth, sensitivity to dietary fats, reduced levels of serum cholesterol, free fatty acids and triglycerides and flattened intestinal microvilli. Scouring severity and incidence was reduced on iso-energetic low-fat diets and growth rates improved by regular parenteral administration of essential lipids. These phenotypes overlap in part with the single reported case of loss of DGAT1 function in humans20, but differ markedly from the normal survival and tolerance of high-fat diets of DGAT1 knockout mice13,21,
Our observations indicate that the mutant cows may provide valuable new insights into the role of DGAT1 in intestinal lipid absorption, postabsorptive fatty acid re-esterification and enteroendocrine peptide release22,23,24,25. In light of ongoing efforts targeting DGAT1 to treat obesity and diabetes26,27,28,29, it will also be interesting to investigate over the longer lifespan of cows any novel effects of DGAT1 deficiency and whether the pleiotropic effects of DGAT1 deficiency observed in mice12 will manifest in a large mammal.
In summary, our findings demonstrate that quality phenotypes for large populations, under long-term trait-specific selective pressure, can be screened to identify new and rare mutations responsible for extreme traits and that these mutations can be employed to establish new animal lines. Combined with genome sequencing and genomic selection for the integration of these traits into the wider population, this approach should result in the rapid identification of animal lines producing foods with improved nutritional qualities. Thus screening of large, well-characterized populations appears to be an approach able to “rescue” new phenotypes generated by spontaneous mutation from the extinction predicted by a mathematical theory of long standing4.
Methods
Population screen
New Zealand's National Herd Testing database records milk volume, protein and fat content and somatic cell count determined by Fourier transformed infrared spectrometry (FTIR, http://www.foss.dk) for 70–80% of dairy cows 3–4 times per season. We retrieved statistical outlier cows with low or high milk fat percentage, high milk protein percentage, or high milk protein : fat content ratio in multiple herd tests over at least two lactation seasons. Candidates were compared to herd mates to exclude cows with extreme milks due to environmental factors and we biased selection towards de novo mutations by excluding candidates with paternal half-sibs or maternal ancestors showing similar, albeit less extreme phenotypes. Candidates were inspected for gross phenotypic abnormalities and their owners interviewed to reveal possible environmental contributions to the extreme phenotypes. The owners were informed of the aims and potential contribution of their animal to the program before the cows were purchased.
Animal procedures
Animals were managed under standard New Zealand seasonal farming routines on mixed ryegrass/white clover pasture and milked twice daily. Adult females were artificially inseminated for calving in July and August. Experimental samples were collected from September to February before cows were dried off in April or May. Lactating females from the founder pedigree were farmed together with unrelated control cows.
In vitro fertilization and embryo implantation into surrogate dams was performed according to routine industry protocols (Artificial Breeding Services, Hamilton, New Zealand). Liver and mammary gland tissues were collected post milking by needle biopsy30, or at slaughter. Samples were snap-frozen in liquid nitrogen and stored at −86°C until use. Animals were monitored after biopsy as described30. Procedures were approved by the AgResearch Ruakura Animal Ethics Committee.
Milk composition analysis
Milk samples were collected during routine morning and evening milking and combined into a representative daily sample. Milk fat and protein content was determined by FTIR. Fatty acid composition was determined by gas chromatography as described31 and casein and whey protein content was determined by HPLC and SDS-PAGE32.
Genetic mapping
The 92 granddaughters of cow 363 indicated in figure 1a (excluding the single cow with 4.16% milk fat content) were genotyped (BovineSNP50 Genotyping Bead Chip, Illumina) by GeneSeek, Inc. (Lincoln, NE, U.S.A.). SNPs with > 5% missing data or minor allele frequency < 5% or that did not map to a chromosome were removed, leaving 39,438 SNPs for analysis. The animals were analyzed for SNP associations utilizing a mixed linear model in Tassel 2.133: milk fat content (in %) = genotype + u + e, where u is a random effect accounting for relatedness and has covariance matrix proportional to the kinship matrix. The kinship matrix was estimated using PLINK34. A Bonferroni correction was used to adjust for multiple testing. The 0.05-level multiple-testing corrected significance threshold is 0.05/39438 = 1.27 × 10−6, which was met by six markers.
Sequence analysis
The sequence of the DGAT1 locus from the founder and one of her low-fat daughters was determined by PCR and Sanger sequencing and compared to sequences from normal cows, to GenBank record AY065621.1 and to bovine reference genome build 4.1, by visual inspection of chromatograms. All sequence variants were confirmed by independent amplification, sequencing and genotyping reactions.
Genotyping of the 8078A>C mutation
Animals from the 363 pedigree, 185 sires frequently used in the New Zealand dairy population and 80 sires and 1595 cows representing a crossbreed herd35, were genotyped for the mutation by the Australian Genome Research Facility using a custom-designed iPLEX™ Gold assay (SEQUENOM) with primers detailed in supplementary file 2.
Identification of splicing regulatory motifs
The effect of the 8078A>C substitution was analyzed against a set of functionally validated hexamer motifs that are statistically overrepresented in exons16,36,37 by querying the RESCUE-ESE web server (http://genes.mit.edu/burgelab/rescue-ese/) with wild-type and mutant bovine DGAT1 sequences.
Analysis of DGAT1 mRNA splicing
Total RNA was prepared from liver samples using RNeasy columns (QIAGEN) and on-column DNAse treatment and reverse transcribed using the SuperScript III First-Strand Synthesis System (Invitrogen). Presence of DGAT1 exon 16 was determined by gel electrophoresis and sequencing of the PCR products obtained with primer pairs to exon 15 and 16, or to exons 15 and 17. Exon 16 skipping was quantified by qPCR on a Lightcycler 480 using the Lightcycler 480 Probes Master System and universal probes (Roche). Briefly, exon16-containing transcripts were PCR-amplified using primers to exons 15 and 16 with fluorescent probe #44 and normalized to DGAT1 transcripts quantified using primers to DGAT1 exons 4 and 5 with probe #98. Detailed parameters are provided in the Supplementary MIQE File. Statistical significance of expression differences was determined using Student's t-test (two-sided, unequal variance).
Yeast expression and diacylglycerol acyltransferase assay
Yeast expression vectors encoding wild-type (Lys232 and Ala232) or mutant (Ala232-ΔE16) DGAT1 were constructed by cloning bovine cDNAs into pYES2 (Invitrogen). Plasmids and vector were transformed into triglyceride-negative strain H124618 and transgene expression was induced using galactose as described (pYES2 Manual, Invitrogen). Twenty OD600 nm of culture was harvested and 1 OD600 nm was retained for mRNA quantification. Cells were sedimented, washed with water and disrupted by vigorous vortexing in 50 μL of 10 mM Tris-HCl (pH7.9), 10 mM MgCl2, 1 mM EDTA, 5% glycerol, 1 mM DTT, 0.3 M ammonium sulfate, Complete protease inhibitor cocktail (Roche), 0.8 mM Pefabloc SC (Roche) and 600 mg of glass beads (diameter 450–600 μm, Sigma-Aldrich). The cell lysate was recovered in 500 μl disruption buffer and cleared by centrifugation at 12000 g-1 for 10 min. Protein concentration was determined using the DC protein assay (Bio-Rad).
Diacylglycerol acyltransferase activity in 50 μg of clarified yeast lysate was determined as described15. TLC plates were exposed to a phosphor imaging screen and scanned (Pharos FX+, Bio-Rad). DGAT1 cDNA was quantified by qPCR using primers to exons 4 and 5 (see above), while endogenous TDH1 transcripts were quantified using fluorescent probe #82 and primers detailed in supplementary file 2. DGAT1 expression levels were computed as the ratio of mean quantification cycle (Cq) for the bovine DGAT1 reaction to the mean Cq for the yeast TDH1 reaction. Statistical significance of gene expression differences was determined using Student's t-test (two-sided, unequal variance).
References
Beja-Pereira, A. et al. Gene-culture coevolution between cattle milk protein genes and human lactase genes. Nat. Genet. 35, 311–313 (2003).
Tishkoff, S. A. et al. Convergent adaptation of human lactase persistence in Africa and Europe. Nat. Genet. 39, 31–40 (2006).
Meuwissen, T. H., Hayes, B. J. & Goddard, M. E. Prediction of total genetic value using genome-wide dense marker maps. Genetics 157, 1819–1829 (2001).
Haldane, J. B. The rate of spontaneous mutation of a human gene. J. Genet. 31, 317–326 (1935).
Kumar, S. & Subramanian, S. Mutation rates in mammalian genomes. Proc. Natl. Acad. Sci. USA 99, 803–808 (2002).
Xue, Y. et al. Human Y chromosome base-substitution mutation rate measured by direct sequencing in a deep-rooting pedigree. Curr. Biol. 15, 1453–1457 (2009).
Roach, J. C. et al. Analysis of genetic inheritance in a family quartet by whole genome sequencing. Science 328, 636–639 (2010).
Bovine Genome Sequencing and Analysis Consortium. The genome sequence of taurine cattle: a window to ruminant biology and evolution. Science 324, 522–528 (2009).
Zimin, A. V. et al. A whole-genome assembly of the domestic cow, Bos taurus. Genome Biol. 10, R42 (2009).
MacGibbon, A. K. H. & Taylor, M. W. Composition and structure of milk lipids. Advanced Dairy Chemistry,. Vol.2. Lipids, 3rd ed., 1–42. Fox, P. F., & McSweeney, P. L. H., eds. Springer, New York, USA, 2003.
Cases, S. et al. Identification of a gene encoding an acyl CoA:diacylglycerol acyltransferase, a key enzyme in triacylglycerol synthesis. Proc. Natl. Acad. Sci. USA 95, 13018–13023 (1998).
Yen, C. L., Stone, S. J., Koliwad, S., Harris, C. & Farese, R. V. Jr. Thematic review series: glycerolipids. DGAT enzymes and triacylglycerol biosynthesis. J. Lipid Res. 49, 2283–2301 (2008).
Smith, S. J. et al. Obesity resistance and multiple mechanisms of triglyceride synthesis in mice lacking Dgat. Nat. Genet. 25, 87–90 (2000).
Grisart, B. et al. Positional candidate cloning of a QTL in dairy cattle: identification of a missense mutation in the bovine DGAT1 gene with major effect on milk yield and composition. Genome Res. 12, 222–231 (2002).
Grisart, B. et al. Genetic and functional confirmation of the causality of the DGAT1 K232A quantitative trait nucleotide in affecting milk yield and composition. Proc. Natl. Acad. Sci. USA 101, 2398–2403 (2004).
Fairbrother, W. G., Yeh, R. F., Sharp, P. A. & Burge, C. B. Predictive identification of exonic splicing enhancers in human genes. Science 297, 1007–1013 (2002).
Cartegni, L., Chew, S. L. & Krainer, A. R. Listening to silence and understanding nonsense: exonic mutations that affect splicing. Nat. Rev. Genet. 3, 285–298 (2002).
Sandager, L. et al. Storage lipid synthesis is non-essential in yeast. J. Biol. Chem. 277, 6478–6482 (2002).
Schennink, A. et al. DGAT1 underlies large genetic variation in milk-fat composition of dairy cows. Anim. Genet. 38, 467–73 (2007).
Haas, J. T. et al. DGAT1 mutation is linked to a congenital diarrheal disorder. J. Clin. Invest. 122, 4680–4684 (2012).
Chen, H. C. et al. Obesity resistance and enhanced glucose metabolism in mice transplanted with white adipose tissue lacking acyl CoA:diacylglycerol acyltransferase 1. J. Clin. Invest. 111, 1715–1722 (2003).
Buhman, K. K. et al. DGAT1 is not essential for intestinal triacylglycerol absorption or chylomicron synthesis. J. Biol. Chem. 277, 25474–25479 (2002).
Cheng, D. et al. Acylation of acylglycerols by acyl coenzyme A:diacylglycerol acyltransferase 1 (DGAT1). Functional importance of DGAT1 in the intestinal fat absorption. J. Biol. Chem. 283, 29802–29811 (2008).
Okawa, M. et al. Role of MGAT2 and DGAT1 in the release of gut peptides after triglyceride ingestion. Biochem. Biophys. Res. Commun. 390, 377–381 (2009).
Lee, B., Fast, A. M., Zhu, J., Cheng, J. X. & Buhman, K. K. Intestine-specific expression of acyl CoA:diacylglycerol acyltransferase 1 reverses resistance to diet-induced hepatic steatosis and obesity in Dgat1-/- mice. J. Lipid. Res. 51, 1770–1780 (2010).
Chen, H. C. & Farese, R. V. Jr. Inhibition of triglyceride synthesis as a treatment strategy for obesity: lessons from DGAT1-deficient mice. Arterioscler. Thromb. Vasc. Biol. 25, 482–486 (2005).
De Vita, R. J. & Pinto, S. Current status of the research and development of diacylglycerol O-acyltransferase 1 (DGAT1) inhibitors. J. Med. Chem. 56, 9820–9825 (2013).
Denison, H. et al. Diacylglycerol acyltransferase 1 inhibition with AZD7687 alters lipid handling and hormone secretion in the gut with intolerable side effects: a randomized clinical trial. Diabetes Obes. Metab. 16, 3334–343 (2014).
Fox, B. M. et al. Discovery of 6-phenylpyrimido[4,5-b][1,4]oxazines as potent and selective acyl CoA:diacylglycerol acyltransferase 1 (DGAT1) inhibitors with in vivo efficacy in rodents. J. Med. Chem. 57, 3464–3483 (2014).
Littlejohn, M. D. et al. Effects of reduced frequency of milk removal on gene expression in the bovine mammary gland. Physiol. Genomics. 41, 21–32 (2010).
MacGibbon, A. K. H. Modified method of fat extraction for solid fat content determination. New Zealand Journal of Dairy Science and Technology 23, 399–403 (1988).
Mackle, T. R., Bryant, A. M., Petch, S. F., Hill, J. P. & Auldist, M. J. Nutritional influences on the composition of milk from cows of different protein phenotypes in New Zealand. J. Dairy Sci. 82, 172–80 (1999).
Bradbury, P. J. et al. TASSEL: software for association mapping of complex traits in diverse samples. Bioinformatics 23, 2633–2635 (2007).
Purcell, S. et al. PLINK: a tool set for whole-genome association and population-based linkage analyses. Am. J. Hum. Genet. 81, 559–575 (2007).
Spelman, R. J., Miller, F., Hooper, J., Thielen, M. & Garrick, D. Experimental design for QTL trial involving New Zealand Friesian and Jersey breeds. Proc. Assoc. Advmt. Anim. Breed. Genet. 14, 393–396 (2001).
Fairbrother, W. G. et al. RESCUE-ESE identifies candidate exonic splicing enhancers in vertebrate exons. Nucl. Acids Res. 32 (WebServer issue), W187–90 (2004).
Yeo, G., Hoon, S., Venkatesh, B. & Burge, C. B. Variation in sequence and organization of splicing regulatory elements in vertebrate genes. Proc. Natl. Acad. Sci. USA 101, 15700–15705 (2004).
Acknowledgements
We are grateful to the Jordan family for the generous donation of cow 363 to the research program. We thank the staff at the Whareroa Research Station for their excellent care of the animal pedigree and for their assistance in milk and biopsy sampling, Sten Stymne (Swedish University of Agricultural Sciences, Alnarp, Sweden) for providing yeast strain H1246 and Howard J Jacob, Dorian J Garrick, Michel Georges and Wouter Coppieters for advice throughout the project. This work was supported by grants VLAC0301 and VLAC0801 from the New Zealand Ministry of Science and Innovation.
Author information
Authors and Affiliations
Contributions
R.G.S. conceived the project. K.L., S.D.B. and R.G.S. designed and directed experiments and analyzed results. A.A.-U., B.H., A.K.H.M., R.J.S. and R.G.S. performed the population screen and Outlier selection. A.A.-U. managed and bred the animal pedigree. Y.v.d.D. and A.K.H.M. analyzed milk fat composition. K.L., H.W., A.A-U., A.B., L.F.A. and N.L.T. contributed to genotyping. E.M.B. and S.R.B. conducted genetic mapping. K.L., H.W., A.B., N.L.T. and C.A.F. contributed to DGAT1 sequencing, bioinformatics analysis, cDNA cloning and expression plasmid construction. P.R. and W.T.J. advised on enzyme function assay. H.W. performed enzyme assays and qPCR reactions. G.V. and S.R.D. provided advice on cow management and animal physiology. K.L., H.W., A.A.-U., A.B., N.L.T. and G.V. performed animal biopsies. K.L., S.D.B., R.J.S. and R.G.S. wrote the first draft of the manuscript and all authors contributed to and read the final version.
Ethics declarations
Competing interests
K.L., H.W., S.D.B., A.A.-U., A.B., E.M.B. N.L.T., C.A.F., S.R.D., R.G.S., L.F.A., Y.v.d.D. and A.K.H.M were employed by ViaLactia Biosciences (NZ) Ltd or its parent company Fonterra Co-operative Group Limited during the research. R.J.S. and B.H. are employees of LIC. S.R.B. and G.V. received fees for professional services from ViaLactia Biosciences (NZ) Ltd. P.R. and W.T.J. received research funding from ViaLactia Biosciences (NZ) Ltd.
Electronic supplementary material
Supplementary Information
Supplementary Dataset 1
Supplementary Information
Supplementary Dataset 1
Supplementary Information
Supplementary Dataset 2
Supplementary Information
Supplementary Dataset 3
Supplementary Information
Supplementary Dataset 4
Rights and permissions
This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License. The images or other third party material in this article are included in the article's Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder in order to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-sa/4.0/
About this article
Cite this article
Lehnert, K., Ward, H., Berry, S. et al. Phenotypic population screen identifies a new mutation in bovine DGAT1 responsible for unsaturated milk fat. Sci Rep 5, 8484 (2015). https://doi.org/10.1038/srep08484
Received:
Accepted:
Published:
DOI: https://doi.org/10.1038/srep08484
This article is cited by
-
Analysis of the impact of DGAT1 p.M435L and p.K232A variants on pre-mRNA splicing in a full-length gene assay
Scientific Reports (2023)
-
A Capra hircus chromosome 19 locus linked to milk production influences mammary conformation
Journal of Animal Science and Biotechnology (2022)
-
A new mechanism for a familiar mutation – bovine DGAT1 K232A modulates gene expression through multi-junction exon splice enhancement
BMC Genomics (2020)
-
Eigen decomposition expedites longitudinal genome-wide association studies for milk production traits in Chinese Holstein
Genetics Selection Evolution (2018)
-
Congenital protein losing enteropathy: an inborn error of lipid metabolism due to DGAT1 mutations
European Journal of Human Genetics (2016)
Comments
By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.