Milk Protein Production

Central Dogma of Molecular Biology

Promoter Intron 1Exon 1 Intron 2Exon 2 Exon 3

AATAAA

DNA

Intron 1Exon 1 Intron 2Exon 2 Exon 3

AATAAA

Unspliced RNA

Exon 1 Exon 2 Exon 3

AATAAA

Spliced mRNA AAAAAAAAAAAA

AAAAAAAAAAAA

Mature ProteinSignal Peptide

Mature Protein

Pre-Protein

Protein

Transcription

Translation

Gene and RNA Structure

Bovine Beta-Casein Gene Part 1TCGAATCCATCTCTATCAATTAATGTAATTCAAAATTGGTGAGAGACAGTCATTAGGAAATTCTCTGTTTATTGCACAATATGTAAAGCATCTTCCTGAGAAAAGGGAAATGTTGAATGGGAAGGACATGCTTTCTTTTGTATTCCTTTTCTCAGAAATCACACTTTTTTGCCTGTGGCCTTGGCAACCAAAAGCTAACACATAAAGAAAGGCATATGAAGTAGCCAAGGCCTTTTCTAGTTATATCTATGACACTGAGTTCATTTCATCATTTATTTTCCTGACTTCCTCCTGGGCCATATGAGCAGTCTTAGAATGAATATTAGCTGAATAATCCAAATGCATAGTAGATGTTGATTTGGGTTTTCTAAGCAATACAAGACTTCTATGACAGTGAGATGTATTACCATCCAACACACATCTCAGCATGATATAAATGTAAGGTATATTGTGAAGAAAAATTATCAATTATGTCAAAGTGCTTACTTTAGAAGATCATCTATCTGTCCCAAAGCTGTGAATATATATATTGAACATAATTAATAGACGAAACAAACCTTGTAAAAATGAGTAGTGTAAAATACAACTACATTTATGAACATCTATCACTAAAGAGGCAAAGAAAGTTGAGGACTGCTTTTGTAAATGGGCTCTTATTAATGAAAAGTACTTTTGAGGTCTGGCTTAGACTCTATTGTAGTACTTATGGTAAGACCCTCCTCTTGTCTGGGCTTTCATTTTCTTTCTTCCTTCCCTCATTTGCCCTTCCATGAATACTAGCTGATAAACATTGACTCACTATAAAAGATATGAGGCCAAACTTGAGCTGTCCATTTTAATAAATCTGTATAAATAATATTTGTTCTACAGAAGTATCTCTAAATAAATGTACTTTCTCTCTTAAAATCCCTCAACAAATCCCCACTATCTAGAGAATAAGATTGACATTCCCTGGAGTCACAGCATGCTTTGTCTGCCATTATCTGACCCCTTTCTCTTTCTCTCTTCTCACCTCCATCTACTCCTTTTTCCTTGCAATACATGACCCAGATTCACTGTTTGATTTGGCTTGCATGTGTGTGTGCTGAGTTGTGTCTCACTCTTGTCAACCCCATGAATGACAGTCCACCAGGCTCCACTATTTCCAGTTAAGAATACTGGAGTGGATTGTGTTTCCTACTTCATTTGATTAATTTAGTGACTTTTTAAATTTTTTTCCATATTCAGGAGGCTATTCTTTCCTTTTAGTCTATACTGTCTTCGCTCTTCAGGTCTAAGCTATCATCATGTGCTTGTTAGCTTGTTTCTTTCTCCATTATAGCATAAACACTAACAACTATTCAGGTTAGCATGAGATTGTGTTCTTTGTGTGGCCTGTGTATTTCTGGTGTGTATTAGAATTTACCCCAAGATCTCAAAGACCCACCGAATACTAAAGAGACCTCATTGTAGTTACAATAATTTGGGGACTGGGCCAAAACTTCCGTGTGTCCCAGCCAAGGTCTGTAGCTACTGGACAATTTAATTTCCTTTATCAGATTGTGAATTATTCCCTTTAAAATGCTCCCCAGAATTTTTGGGGACAGAAAAATAGGAAGAATTCATTTTCTAATCATGCAGATTTCTAGGAATTCAAATCCACTATTGGTTTTATTTCAAACCACAAAATTAGCATGCCATTAAATACTATATATAAACAACCACAAAATCAGATCATTATCCATTCAGCTCCTCCTTCACTTCTTGTCCTCTACTTTGGAAAAAAGGTAAGAATCTCAGATATAATTTCATTGTATCTGCTACTCATCTTTATTTCAGACTAGGTTAAAATGTAGAAAGAACATAATTGCTTAAAATAGATCTTAAAAATAAGGATGTTTAAGATAAAGTTTACAGTATTTTCAGCAAATTTGTTAAAAAATAGAAGCAACTATAAAGATTTGTAACAGTGGTTGCTATTTTCTTTACCACGAGACTAGTTAACAGGCTGTATTAAAAGATCTTTTCTTGAATTAAATATTTTCAATTTGATTAAACATACCTCAGCCATAAAGGCAAGCACATTTAATTTATACTATGGGAATTTGAATAATTGTTACTGAAGAAGCTCTACCAACAAAAAGTTTATAGAGCTAGCATATTTAGTCAAGAGATAAAGAGGGTTGTTAGGATACATGTGCTATTTGAAAGGTATTTATAAAAGAAGAGTATATTTATTAAAATTGCTCAGAACATCCAAATTTCAAGTTTATCATTTATCTTACAATATTTCAAAAATATTAAAATAGATACATGAAATACAGAAGTAAATTAAAGAGAAAGTATTTTATTTTGTAAAAAAAAATTCTAGGTTGGACAGGGAGTACCAGGAAACAAAAAACAATGAAAAATGTGATCTGACAGAAATTATAGCTCAAAGTATAGTAGTCAGTAATGAAATGGCTTAAAAATTGGCATATAAAATGCTAATTATAAAATAAACAAAATGTAATAATACCCTCCCTACATGTAATGAACTCTGAGTATTATACTCTTTTTTGAAGTCTTGACAATGAAAATTTATTTAGACTTTTATAGACATCTTGGATAAAGTAAAACAAATTACGAATTAGCATCCATGAGAAAAATATAGAAAAATTTCTTAATGTAGTTTGCAAATCTGGGGATTGAAGATGTGTGTCAAGAGATTGTGATGGCAGACATTTTTTTTCAGACTATAAAATGCACAAACAACCATTTAATACATTTTGGTCAAAAATAGTATGTATTTTATTTTATGCTACAGGAGAGTAGTCTAAAGTAGGACTGGGCAGAGATCTGACACCCTGGTAATCACCGAGAGATAGTACACAGTCTCTGTAGAGAAAATAAGCATAGTGTATGATCTCTAAAATTATGTGGACAAAGGGGAGATAACATTAGGCATGTGGGGATGAAGACTGAGTACAGAAGAACAATCTAGTCAGTCCAAGAAAACATGTGGATCAATGGAACAAATAGAAGAAATGCTAAAATGAAACAGAAGTCTTACTGGAAATAAAAGATATGAGGAAGACAAACATTCATGAAAATCACTTAGTTTAGTAGAGAAAAGATAAAAATAAAGTATTACCTTCTTCTTCATATACATTGTTTGATCAGATGCCCCTCAATAAAACTGAGTCTCCAACAGAACTGAAACTTTAATATTTTGTTCACTGCTCTAATCCCAGAATCTAAGACATATCTGGCAATAAAAATTAATAAATAAATATTTTTAATAAGTAAATCAATCACTTAATTTTTCTGTAAGTATCTGTAACTTCTCTTCTGTCTTTCCAAAAAACACTCATAAGTACTGTGAATAAGATGAAAAGAGTGAAATAAGATATAGGCTGTTAGCTGAAAACATCTGGATGGCTGGCAGTGAAACATTAACTTGAAATGTAAGATTAATGAGTAATAGTAAATTTTAACCTTGGCCGTATGATAAAATGTCTATTAATATTTTTCTAAAATACAGGGCTTTTTGTTTTTGCCATGAGGTTTGCAGGATCTTGGTTCCCTGATGAGGGATCAAACCTGGGCTCCCCTGGAAGCACGGAGTCTTAGATATTTGTATTATACACTATCTTTGGTTTCTTTTAAAGGGAAGTAATTCTACTTAAATAAGAAAATAGATTGACAAGTAATACACTATTTCCTCATCTTCCCATTCCCAGGAATTGAGAGCCATGAAGGTCCTCATCCTTGCCTGCCTGGTGGCTCTGGCCCTTGCAAGAGAGGTAAATACAGAAAAAATGTTGAAATAAATAAGACTAGTACTATCTGCTATGTGTAGAAAATTCATTACCAACATTGTAAATGTATAAATAATGCACAATCTCAGATTTTTTTTGAATGCTAAGAAAGTCATTTACGTTCATCCACTATCTCAGTAGTATCCTATGGGACCACAAGTCTGAGTCTAGTGCTTTCTATAGTATTGTACCATCTGTACCATCAATCCCTAAAGAAAAAAGAAAATAAACCAATAAGCAACAGACTAACAAGAAGGAACACAGATAAGAACAAAAAGTGAGTAATATTGCATAAATACAATTGCATGCATATACAATCTAGATAAATATATCTTATTCCAGTGATGAAATATTTGTATCCCTTACTGTAGAGTGCTAGGTTTAGCTGTGTCTATTCAACACAGGATGATACTCCAGAGGATGGTATATCAGACAACAATAATAAATATGTTCATAATTATAATAAAAAGTGTTCAGTAAAAATTAAAATAACTCCTTTTCTGTTACCCATAAAAACTCTTCATTAAAGTAAAACAAAAATATACTAATGAAAGTTACTAAATTTAAAAGACTCTCAAAAGACATATAACATTTTTATTTTTCAGATTTGTGAAATAGATAGCTCTGAATAAAGCAAGTAAAAATTAGGTAGGAAAATATTTAATAATGAGTTGACTGTGGGAACTAAAGTGTTTTTTTTTCTCTTTAGCTGGAAGAACTCAATGTACCTGGTGAGGTAAGATATTTTTATACAAAGAAAAAAATTAATTTAACTGTAAAATAGTAACAGTCTCTAATGATCTGGCAGAAGACTCAGCTAATTGTCAATTTTTATTTTTCCTTTATAGATTGTGGAAAGCCTTTCAAGCAGTGAGGTAAGATAGTGTTCATTCAGAGGCAATTTCCCAAATTTAGAGCAATAAAATGCTGTATTATCTTTTTGTGTTAC

Bovine Beta-Casein Gene Part 2ATTAATGGCAACCCACTCCAGTATTCTTGCCTGGAAAATCCCATAGAGGAGGAGCCTGGTAGGCTGCAGTCCACGGGGTCGCTAAGAGTCGGACAGGACTGAGCGACTTCCCTTTCACTTTTCACTTTCATGCCTTGGAGAAGGAAATGGCAACCCACTCCAGTTTTCTTGCCGGGAGAATCCCAGGGACGGGGGAGCCTGGTGGGCTGCCGTCTATGGGGGTCGCACAGAGTTGGACACGACTGAAGCGACTTAGCAGTAGCAGCAGCAGGACAGTTAAGGTTTCTCTAATAGCTCAGTTGGTAAAGAATCTGCCTGCAGTGCAGGAGGACCCTGGGTTCGATTCCTCAAGATCTGCAGGAGAAGGGATAGGCTACCCATTCCAGTGTTCTTTAAGCCAATGTGGCTATGTACTGACGGGTAACTATTGTCAATTTCCACTCTGTATATTTAAAGGAATAAATGTGTAGAAGGTTTAATATTCTAGTAATTTCTAAATGGGTTTGTTATTTGAAATTGTGTCATTGTGCCCTGCTTTTTTTCCTTAATGAACTGTACAGTCCTCTTTTCTGTCTTGAACTTTCTATGTTAACCTCCTTCCATGCTTTGGCATTTATTGAGCACTTTCTGTTTCAGACTTTGACTAGGAACTGCAGTACAAACTAGAAAGAGGGATGCCCTTTGTAAAGTGTGAGCAGACATGCAAATGGACATATTTTATTATTATACAAGCAATCCAGTACACAAGAGGCAGTGAGAATGAGTGTAGTCCTAAATCTGCCTGGTGGAATGAGGTAGATAATAACCCTATGCCACTCTTTTTGGCTCTGTCATCAGCTGTTGGTTAAATAGTGCATCAATATACTTTGTCTCTTCACAAAGGTCAAAAGATGCTGCTGCTGCTGCTAAGTCGCTTCAGTCATGTCCGACTCTGTGCGACCTCATAGATGGCAGCCTGCCAGGCTCCCCCATCCCTGGGATTCTCCAGGCAAGAATACTGGAGTGGGTTGCCATTTCCTATTCCAATGCATAAAAGTGAAAAGTGAAAGTGAAGTCGCTCAGTCGTGTCCGACTCCTAGTGACCCCGTGGACTGCAGCCTACCAGGCTCCTCCATCCATGGGATTTTCCAGGCAAGAGTACTGGAGTGGGTTGCCATTGCCATCTCCAAAGATCCTTATAGGAGGGTATGTATTTCTTATAATTCATTAGAAGCCTAAACATAACCAGGGAACTAAGAATGATTAACAAGTTCATGCGTGGTTATTATTATATATTTTCAATGACTATTATTCTTTTAGAACCAGATGAAAATATTAGAGATCATTTGTTGTCCTAAGGAGAGAACAGGATGATTGAGAGACATGTATGCATGCAAAGTTACTTCAGCCCTTTCCAACTCTGTATGACCCTATGGACTGTAACCTGCCAGGTTCCTCTGTCTATGGGATTCTCCAGGCAAGAATACAGGAGTGGGTTGTCATCTCCTCCAGGGCCTAGGAATTGACCTGCATCTCTTACGTCTCCTGCATTGGCAGGCAGGTTCTTTACCACTAGCACCACCTGGAAGCCCGATTAGTATCTGTTAAATGCCTCTTTGAGTACTATGCTCTCTAATCCTCTTTTTCTTGATTGCATCATCTTTCTTTTTATACAACAGCCTTATTCAGAAGAGTGGAACATAAACTTTCAGCCATAAAATAATGATATTATCAAATGAGCTGTCCATATTAATCTATTAAATTTCTTCATTTTCTGATTTATGTTGACAAATAAGAATTTTTTTTAAAGCTAGACCTGATTTTATTTTTATTTTTCCAAAGGAATCTATTACACGCATCAATAAGGTAAAACCCCTCATATTTAAATGTACATTTTTTTAAATTTCATGTTTGATTTTTATAAACAGCATTTCTTTATGTATTTTTTTTTTAACCAGAAAATTGAGAAGTTTCAGAGTGAGGAACAGCAGCAAACAGAGGTAATTTGTTCACTATGAGTATATTTTGAGAAGTATTATGAAACATAACACATAAAAAGATTTATAATAATTATGTTCAGTCTAAGAATGGTAATATAAATGTCAGTGCAAGAAATAAAAACTTTGACAAAATGAAAATATTTTAAAAATATAAACGCATTTTAAAACACATAATCAAATTTCACAGTATAGAATAAATAGCTAAGAATAATTATTGATGTATTCATTTTACTAATGGTATACCTGGTTTTAATAACTGCATATTAGTAGGAACATTTCCAGACTAGGGACTGTGATCCCCTTATTCTAATGATGGATATGCTGATGAAAGACAGTAGGGTGACAGTGTGGCACTAATCCTCATCTGATCATTTATCAGCTGTATAACCTTGGCTCCATGTTTCTCTGTACATCATTTTCTTCACCTGTAAATTGAGAATATTCATAATTACCCAGAGTTGATGAACTGACACACAATGAATATTCAGTGGTTTTATATTATATTTGATAGCTTTTATACTCACATTTATGGATGTGTGGAGTTCTAAAAAGTATTTCCATTGCCCAGATGAGAGAAGTGAGGTACAGGACAATTGAGTATGCAAATGTGTGACCATACCACATAGTTATTAAATAGCAGAACTTGCTTAAAAACAAGGATTTGCGGACAATGTAAAATTCTTTCATTATATTACTCTTGTGGTAACATATTTATCTAATTATGATATTTAAGCTTTCCTCTTTTATAATTGAAGTTTGATTGTTTGGCACTTAGGCCAAATTCTAAATCAAAATGAATTTACAACTTGATGCCTTTGAAGACTCAAGATTACCACCTTCTACCAAGAGAAGTAGTGCTAGAAGTTGGCCATTGTTAAGGAACTCCTTGAATTAAAAAAACACATATTAAGACTTAGTTTTCATTAAAACAAACAAAAATAAACCTCAGAGTAACTTTTAAAGTCTTTTTAAAATGGATCTTTCTTTGTTATATGAAACCAGTTTGGACTATTATCCAAAGTATGTAGCTACCACTCTGCAGGAACTCAGGAAGAGGTGGAATAAGTGTTGAAATCTCCAAACCCTGATTTCACTTGACTCTCTGATTTCACCTGTGAAGAAAGTGGGTTAATGAGAAATCCTTCAGTGAGCATTTTACTCATTAGTCTTCATATGACCCCAATTTCTTAACCAAACCAAATGGAAGATTTTCTTTCTCTCTCTTCACTGAATTATGTTTTAAAAAGAGGAGGATAATTCATCATGAATAACAATTATAACTGGATTATGGACTCAAAGATTTGTTTTCCTTCTTTCCAGGATGAACTCCAGGATAAAATCCACCCCTTTGCCCAGACACAGTCTCTAGTCTATCCCTTCCCTGGACCCATCCATAACAGCCTCCCACAAAACATCCCTCCTCTTACTCAAACCCCTGTGGTGGTGCCGCCTTTCCTTCAGCCTGAAGTAATGGGAGTCTCCAAAGTGAAGGAGGCTATGGCTCCTAAGCACAAAGAAATGCCCTTCCCTAAATATCCAGTTGAGCCCTTTACTGAAAGCCAGAGCCTGACTCTCACTGATGTTGAAAATCTGCACCTTCCTCTGCCTCTGCTCCAGTCTTGGATGCACCAGCCTCACCAGCCTCTTCCTCCAACTGTCATGTTTCCTCCTCAGTCCGTGCTGTCCCTTTCTCAGTCCAAAGTCCTGCCTGTTCCCCAGAAAGCAGTGCCCTATCCCCAGAGAGATATGCCCATTCAGGCCTTTCTGCTGTACCAGGAGCCTGTACTCGGTCCTGTCCGGGGACCCTTCCCTATTATTGTAAGTCTAAATTTACTAACTGTGCCTGTTTAACTTCTGATGTTTGTATGATATTCGAGTAATTAAGAGTCCTATAAAAAAATGAATAATGAATGGTTCCAAAATAAGCATAGCTGAGATTAATGATTGTCAGCATTAGTTATAAATAGAATAAGCTGGAGAACCTTCACCTCCCCTCCACCACCAGATCTCAATGTCTAGGCTTACCCGTGGAGATTCTGATGTAATTGTTCTTTCTATGTAGAAGAAACTTATTGGGAAGAAATAATATAATGGACTATGATTTAATTGGTCTGTTGAGAACCAATTAAATTAGATGAAAGCGATTAAGTACAATAAAGCCAAAATTGAATTTGATAATCTCATTTGGCTAAGAATAACAAACCTAAGAAGGTTTGCTATTTTCTACAATTTTGAAGTTCTCCTTATGCACAATTATTTCACCACATGACTCATTTCACATCGTGTTTTTGATATATGAGCATATGAGGGAAAAATACTGAGATGCTTATTTCAATACTCAGGGAAAATTTATTGCCAAAAGGCAAGAAATGTATAATTCATTCACTTATTTTATTTTATTATTTTTTTTATTTTTAAGGTCTAAGAGGATTTCAAAGTGAATGCCCCCTCCTCACTTTTGGTAAGCTTTAGGATATTGGAGGCAGACTGATCATTTTTATAGTTAATATCTTTTACATTTCATTTTCCTGGATAAGCCCCAATAGTAGCAATTTCCATCAGTGTACCAGCTTAAAGATTAATTATAAATTTATTTTCAATGATTGACTGTTATTTACTGGCCTGAAATTATGTATCTGTTATATTTCAAATAATGCAAAACTGTATATATATGGTGTTTACAGATTTGATTGGTTTTCTTTCAATAGCCTATATCCTTATTATTGATTGTCATCATTTATAGAAAAAACTGAAAATAATTTCTTATACTTTTATGTAAACCTGTTAGAGCTTATTTTAAAGATCAACTGCATTCACATTTCTAATCTAGTCATTATGAGCTTCAATAGTTTTATCTCACTTAAAATATATATATTGTCTTTTAATTCATGAGTCAAAATACAATCTCACAGTCCAGATATGGGACTTAAAAGGGGGATAGAATATAGTTTTGATATTCTTAACAATACACATCCTTTTGTGATCATGATTCAGCAGACATTTAATAAAATGATTCCAAGTAAGCCGATGTTTGGTCCTAGAGGAATTTTTATAACCTTTAAGAGAAGGCATAGCATGGTGTTTTTGTAATAAGATTTCTTTTATGAAAAAGTCACACCAAAATTGCAAATGGGGGTGAGATGAAGAGTTATAACATATAACTAAATCTATGTTTGTTCTCTATTCCACAGAATTGACTGCGACTGGAAATATGGCAACTTTTCAATCCTTGCATCATGTTACTAAGATAATTTTTAAATGAGTATACATGGAACAAAAAATGAAACTTTATTCCTTTATTTATTTTATGCTTTTTCATCTTAATTTGAATTTGAGTCATAAACTATATATTTCAAAATTTTAATTCAACATTAGCATAAAAGTTCAATTTTAACTTGGAAATATCATGAACATATCAAAATATGTATAAAAATAATTTCTGGAATTGTGATTATTATTTCTTTAAGAATCTATTTCCTAACCAGTCATTTCAATAAATTAATCCTTAGGCATATTTAAGTTTTCTTGTCTTTATTATATTTTTTTTAATGAAATTGGTCTCTTTATTGTTAACTTAAATTTATCTTTGATGTTAAAAAGAGCTGTGGAAAATTAAAATTGGATAGAATTC

RNA Processing

Binding of General Transcription Factors to DNA

Binding of RNA Polymerase II to the Transcription Complex

Controlling mRNA Transcription

Formation of a Stable Initiation Complex

Initiation of Transcription

RNA Transcription

mRNA Capping

mRNA Polyadenylation

mRNA Splicing

Final Structure of Milk Protein mRNA’s

Milk Protein mRNA Degradation

Milk Protein mRNA Stabilization Factors

Cortisol and Prolactin induce production of mRNA stabilization factorsto prevent degradation by nucleases.

Control of Milk Protein Transcription

Inhibiting Milk Protein Gene Transcription

Milk Protein Transcription Inhibition

Progesterone induces LIF production, so LIF is involved in the progesterone inhibition of lactation

mRNA Destablization

Model for Milk Protein Transcription During Pregnancy

Model for Milk Protein Transcription During Lactation

Milk Protein Translation

Ribosome Structure

Initiation of Protein Synthesis

Identification of the Initiation Codon

Protein Translation

Termination of Protein Synthesis

Milk Protein Production

Signal Peptide Function

Signal Peptide Cleavage

Completion of Protein Synthesis

Glycosylation of Milk Proteins

Glycosylation of Milk Proteins

Phosphorylation of the Caseins

Regulation of Protein Synthesis

Mammary Epithelial Cell Cartoon

Transgenic Animals

• Animals that contain DNA (Genes) that they would not obtain through normal breeding or mating practices.

• Genes can be introduced across breed, species or kingdom.

• The gene is stable once incorporated into the animal.• The gene is passed on to the animals offspring in a

normal fashion

Transgenic Animals

DNA Microinjection into the Embryo

Gene Knock-out Mice

Gene Knock-out Mice

Gene Knock-out Mice