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Bovenhuis and Schrooten 2002

ISOLATION, LINKAGE MAPPING AND ASSOCIATION ANALYSIS WITH MILK PRODUCTION TRAITS IN MANCHEGA SHEEP BREED OF THE OVINE ALPHA-AM YLASE GENE CLUSTER. J.H. Calvo 1 , S. Marcos 1 , A.E. Beattie 2 , C. González 1 , J.J. Jurado 1 , M. Serrano 1

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Bovenhuis and Schrooten 2002

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  1. ISOLATION, LINKAGE MAPPING AND ASSOCIATION ANALYSISWITH MILK PRODUCTIONTRAITS IN MANCHEGA SHEEP BREED OF THE OVINE ALPHA-AMYLASE GENE CLUSTER. J.H. Calvo1, S. Marcos1, A.E. Beattie2, C. González1, J.J. Jurado1, M. Serrano1 1Departamento de Mejora Genética Animal, INIA, 28040 Madrid, Spain 2AgResearch Molecular Biology Unit, Department of Biochemistry and Centre for Gene Research, University of Ottago, Dunedin (New Zealand)

  2. Introduction 3.7 cM (Beever et al. 1997) Bovenhuis and Schrooten 2002

  3. AMY2B AMY2A AMY1A AMY1B P-AMY AMY1C n Introduction a- Amylase Funtion: Catalyzes the endohydrolysis of a1-4-glucosidic linkages in oligosaccharides and polysaccharides Human AMY cluster Groot et al., 1989

  4. Bovine Human Sheep breeds: Exons 4 and 5 Exon 2 1Manchega white variety 1 Manchega black variety 1 Rasa aragonesa 1 Assaf 1 Awassi Material and Methods Genomic walking and genetic polymorphisms Primers Samples Mouflon

  5. Material and Methods PCR-SSCPs • PCR conditions: standard • 7% polyacrylamide gel (acrylamide-bisacrylamide (49:1), and 8.7% glycerol) • Running conditions: 8W, 20ºC for 12 hours • DNA bands were visualized by silver staining Linkage mapping • International Mapping Flock (IMF) pedigrees

  6. Material and Methods Association studies Daughter design • 13 sire families (29 daughters per sire) • Estimated breeding values (EBVs) of standardized milk yield and fat and protein contents Model: • Across families analysis Yij= m + Fi + Gj + (FxG)ij + eij • Withinfamily analysis Yi= m + Gj + ei

  7. 198 bp 231 bp 389 bp In4 Ex 4 Ex 5 Results and Discussion Isoform 1 No polymorphism were found

  8. Insertion/deletion 3 b.p. SNP SINEbov 195 bp 198 bp 389 bp 361 bp 1070 bp 147 bp In4 Ex 3 Ex 4 Ex 5 Results and Discussion Isoform 2 20 bp Ex 2

  9. Pro Asn Glu Asn Val Val Ile Pro Asn Glu Asn Val Ile Thr Results and Discussion PCR-SSCPs -/- Val/Val Val/-

  10. Bovine Amy Human Amy2B Human Amy2A Human Amy1A DNA / protein DNA / protein DNA / protein DNA / protein Ovine Amy isoform 1 95 (672) / 83 – 96* (139) 90 (428)/ 88 – 94* (136) 88 (428)/ 87 – 92* (136) 88 (428)/ 87 – 91* (136) Ovine Amy isoform 2 87 (676 ) / 84 - 90* (139) 89 (618)/ 88 – 95* (180) 88 (618)/ 88 – 94* (180) 88 (618)/ 88 – 94* (180) Results and Discussion Identity percentage * Amino-acid conservative identity. Base-pair and amino-acid overlap are presented within parentheses.

  11. INRA130 S29 INRA003 AMY BM4129 MCM58 BL41 10 cM AE57 BMS482 Sheep chromosome 1 Results and Discussion Linkage mapping

  12. Results and Discussion Association studies Within Trait Average EBVs Statistics Family Val/Val -/- F P 6 Milk yield 13.09 2.42 7.440.0172* Fat content 0.08 0.00 0.98 0.3402 Protein content 0.21 0.20 0.01 0.9262 Across Families Milk yield 19.04 20.7 0.61 0.4354 Fat content 0.04 0.08 1.14 0.2886 Protein content 0.02 0.04 0.58 0.4465

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