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Introduction Aims and Objective Significance Materials and Methods Protein Sequence of Livestock Species Protein Sequence of Orthologous Species Natural Variants Post Translational Modifications

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Protein Structure Prediction Protein Mutation Comparison Conservation Scenario of cow β-casein in 41

species Identification of closely related species to cow Gene Structure Analysis Identification Of Novel Mutational Sites, Protein

Quality And Composition Conclusion Recommendation

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Caseins are the major protein fraction and their function is totransport calcium phosphate in milk.

β-casein is a fraction which forms up to 45 % of bovine caseins in milk.

Milk protein variants are also a useful tool for breed characterization, diversity, and phylogenetic studies.

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Literature review on milk proteins with a particularattention to recent findings on milk protein geneticvariants.

To study the effects of different protein variants on milk yield and composition.

To understand the biological significance of the Post Translational Modifications and genetic variations.

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β-casein milk protein was studied in 41 species.

12 closely related species to cow were examined in detail.

Critical residues were predicted which can play vital role in improving milk quality.

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3

2

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β-casein protein sequence of livestock species have 222-224 base pairs length.

Artiodactyla

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Sequence lengthfor β-casein proteinvaries between188-302 base pairin 41 species.

Species of orderDiprotodontia havemaximumsequence length.

Species of sameorder consisted ofrelatively closersequence length.

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Cow has 12 Natural Variants A1, A2, A3, B, C, D, E, F, G, H1, H2 and I.

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Glycosylation

Phosphorylation

Post Translational Modifications

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2D Protein Structure

>CowMKVLILACLVALALARELEELNVPGEIVESLSSSEESITRINKKIEKFQSEEQQQTEDEL

3D Protein Structure

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Template Search

BLAST

Model Building

Model Refinement

2D Protein Structure 3D Protein Structure

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3D Protein Structure2D Protein Structure

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2D Protein Structure 3D Protein Structure

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2D Protein Structure 3D Protein Structure

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To check residue variations in β-casein of 41 species logo was generated using skylign.

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Multiple sequence alignment of β casein protein of 41 species

Natural Variants, Glycosylation sites and Phosphorylation sites have been observed

Species of each order contains mostly conserved

Phosphorylation sites within the same order

No glycosylation site was found in livestock species

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Multiple sequence alignment of β-casein protein sequence in 41 species.

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Pairwise Alignment

Check Identity % of β-casein

of cow with that of other species

Identify closely species to cow on the basis of

identity %

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Exons number was retrieved for gene structure analysis

Species of order Artiodactyla have 9 exons except sheep with 7 exons.

Primates’ species possess

6-7 exons.

Species as dog, mouse, rat,

horse, donkey, rabbit and pikahave 9 exons.

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Milk quality of Cow and Buffalo is closely

similar because of mostly conserved variations in those

species.

Phosphorylated sites play important role in

milk quality.

Highlighted Post translational

modifications can be introduced in other species than cow to enhance their milk

quality and milk composition.

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The relationship between polymorphic variants of β-casein and the composition of milk can be studied in further research

Dolphin can be considered as an important species for future work on milk quality and milk composition due to 70% identity of β-casein milk protein of dolphin with that of cow and conserved Phosphorylation sites with primates species.

Microbat can be considered for further research because Phosphorylation sites predicted in microbat are conserved in order Artiodactyla.

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Worthy Director, CIIT SWL, Dr. Abdul Waheed.

Head of Department of BioSciences, Dr. Shazia Mannan.

Supervisor, Ms. Samina Kausar. Co-supervisor, Mr. Muhammad Asif All faculty members.

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Caroli A.M., Chessa S. and Erhardt G.J.: Milk protein polymorphisms in cattle: effect on animal breeding and human nutrition. Dairy Science. 2009, 92: 5335–52.

Jensen HB, Holland JW, Poulsen NA and Larsen LB: Milk protein variants and isoformsidentified in bovine milk representing extremes in coagulation properties. Dairy Sci.2012, 95:2891–2903.

Ceriotti, G., D. Marletta, A. Caroli, and G. Erhardt.: Milk protein loci in taurine (Bos taurus) and zebu (Bos indicus) populations bred in hot climate. Anim. Breed. Genet. 2004a, 121:404–415.

Blom, N., Gammeltoft, S. and Brunak, S: Sequence and structure-based prediction of eukaryotic protein phosphorylation sites. Molecular Biology. 1999, 294(5): 1351-1362.

Jensen HB, Holland JW, Poulsen NA and Larsen LB: Milk protein variants and isoformsidentified in bovine milk representing extremes in coagulation properties. Dairy Sci.2012, 95:2891–2903.

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