Introduction to Animal Biotechnology

Introduction to Animal Biotechnology

Supamit Mekchay

Department of Animal and Aquatic Sciences

Faculty of Agriculture, Chiang Mai University

Introduction

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Animal Production

Introduction

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People in the world:

Introduction

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- 6 billion people - in 2050, it will be probably double

Introduction

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Introduction

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Biotechnology is applied in animals to enhance effecience of food production and improve health for animals and human.

Introduction

What is Biotechnology ? It is the manipulation of living organisms or parts of organisms to make products useful to human

It is using knowledge of cell to modify their activities in order to make living organisms more effective in serving people

Biotechnology is a broad discipline in which biological processes, organisms, cells or cellular components are exploited to develop new technologies

(Burgess, 2017. Nature Reviews Genetics 274, doi:10.1038/nrg.2017.30) AgBiotech - CMU

Molecular Breeding

Transgenic animals

Animal Biotechnology

Food (meat, milk, egg) Environment

Reproduction

Health (Human, animals)

Recombinant DNA/Protein

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1. อาหาร

Introduction

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Basic of Animal Biotechnology

Supamit Mekchay



Basic of animal biotechnology

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DNA


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The Nobel Prize in 1993

PCR-Polymerase chain reaction

Kary B. Mullis

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Polymerase chain reaction (PCR)

Denaturing DNA sample to separate DNA (94 oC, 30 sec)

(72 oC, 5 min)

(10 oC, 99 h)

Annealing Primer bind to DNA strands (55-60 oC, 30 sec)

Elongation Polymerase synthesizes new DNA strands (70-72 oC, 30 sec)

Denature DNA sample to separate DNA (94 oC, 3 min)

32 X

32 X

95 oC 30 sec

95 oC 3 min

55-60 oC 30 sec

72 oC 60 sec

72 oC 5 min

10 oC 99h


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A B C

ACTT

ACTT ACGT

ACTT ACGT

ACGT


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Molecular DNA marker testing

AACGCTGATCGAACGGTCGTCGACGTCGTCGTGCTGAAGGACTCGTAGGCCACGTGGCA

TTGCGTCTAGCTTGCCAGCAGCTGCAGCAGCACGACTTCCTGAGCATCCGGTGCACCGT

AACGCTGATCGAACGGTCGTCGACGTCGTCGTGCTGAAGTACTCGTAGGCCACGTGGCA

TTGCGTCTAGCTTGCCAGCAGCTGCAGCAGCACGACTTCATGAGCATCCGGTGCACCGT

X RsaI

RsaI

RsaI = GTAC

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CT

RYR

SSC6

CT CT CC TT CT CT CT CT CC TT CT CC TT CC CT TT CC

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automated DNA sequencer


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>KR865958.1 Sus scrofa MC1R gene, complete cds

ATGCCTGTGCTTGGCCCGGAGAGGAGGCTGCTGGCTTCCCTCAGCT

CCGCGCCCCCAGCCGCCCCCCGCCTCGGGCTGGCGCCAACCAGACC

AACCAGACGGGCCCCCAGTGCCTGGAGGTGTCCATTCCCGACGGGC

TCTTCCTCAGCCTGGGGCTGGTGAGCCTCGTGGAGAACGTGCTGGT

GGTGGCCGCCATCGCCAAGAACCGCAACCTGCACTCGCCCATGTAC

TACTTCGTCTGCTGCCTGGCCGTGTCGGACCTGCTGGTGAGCGTGA

GCAACGTGCTGGAGACGGCCGTGCTGCTGCTGCTGGAGGCGGGCGC

CCTGGCCGCCCAGGCCGCCGTGGTGCARCAGCTGGACAATGTCATG

GACGTGCTCATCTGCGGCTCCATGGTGTCCAGCCTCTGCTTCCTGG

GCGCCATCGCCGTGGACCGCTACGTGTCCATCTTCTACGCGCTGCG

CTACCACAGCATCGTGACGCTGCCCCGCGCGGGGCGGGCCATCGCG

GCCATCTGGGCGGGCAGCGTGCTCTCCAGCACCCTCTTCATCGCCT

ACTACCACCACACGGCCGTCCTGCTGGGCCTCGTCAGCTTCTTCGT

GGCCATGCTGGCGCTCATGGCGGTACTGTACGTCCACATGCTGGCC

CGGGCCTGCCAGCACGGCCGGCACATCGCCCGGCTCCACAAGACGC

AGCACCCCACCCGCCAGGGCTGCGGCCTCAAGGGCGCGGCCACCCT

CACCATCCTGCTGGGCGTCTTCCTCCTCTGCTGGGCACCCTTCTTC

CTGCACCTCTCCCTCGTCGTCCTCTGCCCCCAGCACCCCACCTGCG

GCTGCGTCTTCAAGAACGTCAACCTCTTTCTGGCCCTCGTCATCTG

CAACTCCATCGTGGACCCCCTCATCTACGCCTTCCGCAGCCAGGAG

CTCCGCAAGACCCTCCAGGAGGTGCTGCAGTGCTCCTGGTGA

mRNA sequence


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>AQA28405.1 melanocortin 1 receptor

[Sus scrofa]

MPVLGPERRLLASLSSAPPAAPRLGLAANQTNQTGPQCLEVSIP

DGLFLSLGLVSLVENVLVVAAIAKNRNLHSPMYYFVCCLAVSDL

LVSVSNVLETAVLLLLEAGALAAQAAVVQQLDNVMDVLICGSMV

SSLCFLGAIAVDRYVSIFYALRYHSIVTLPRAGRAIAAIWAGSV

LSSTLFIAYYHHTAVLLGLVSFFVAMLALMAVLYVHMLARACQH

GRHIARLHKTQHPTRQGCGLKGAATLTILLGVFLLCWAPFFLHL

SLVVLCPQHPTCGCVFKNVNLFLALVICNSIVDPLIYAFRSQEL

RKTLQEVLQCSW

Amino acid sequence


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BLASTN alingment (nucleotide)

Query 1 ATGCCTGTGCTTGGCCCGGAGAGGAGGCTGCTGGCTTCCCTCAGCTCCGCGCCCCCAGCC 60

||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||

Sbjct 1 ATGCCTGTGCTTGGCCCGGAGAGGAGGCTGCTGGCTTCCCTCAGCTCCGCGCCCCCAGCC 60

Query 61 GCCCCCCGCCTCGGGCTGG-CGCCAACCAGACCAACCAGACGGGCCCCCAGTGCCTGGAG 119

||||||||||||||||||| ||||||||||||||||||||||||||||||||||||||||

Sbjct 61 GCCCCCCGCCTCGGGCTGGCCGCCAACCAGACCAACCAGACGGGCCCCCAGTGCCTGGAG 120

Query 120 GTGTCCATTCCCGACGGGCTCTTCCTCAGCCTGGGGCTGGTGAGCCTCGTGGAGAACGTG 179

||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||

Sbjct 121 GTGTCCATTCCCGACGGGCTCTTCCTCAGCCTGGGGCTGGTGAGCCTCGTGGAGAACGTG 180


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BLASTP alingment (protein)

Query 1 MPVLGPERRLLASLSSAPPAAPRLGLAANQTNQTGPQCLEVSIPDGLFLSLGLVSLVENV 60

MPVLGPERRLLASLSSAPPAAPRLGLAANQTNQTGPQCLEVSIPDGLFLSLGLVSLVENV

Sbjct 1 MPVLGPERRLLASLSSAPPAAPRLGLAANQTNQTGPQCLEVSIPDGLFLSLGLVSLVENV 60

Query 61 LVVAAIAKNRNLHSPMYYFVCCLAVSDLLVSVSNVLETAVLLLLEAGALAAQAAVVQQLD 120

LVVAAIAKNRNLHSPMYYFVCCLAVSDLLVSVSNVLETAVLLLLEAGALAAQAAVVQQLD

Sbjct 61 LVVAAIAKNRNLHSPMYYFVCCLAVSDLLVSVSNVLETAVLLLLEAGALAAQAAVVQQLD 120

Query 121 NVMDVLICGSMVSSLCFLGAIAVDRYVSIFYALRYHSIVTLPRAGRAIAAIWAGSVLSST 180

NVMDVLICGSMVSSLCFLGAIAVDRYVSIFYALRYHSIVTLPRAGRAIAAIWAGSVLSST

Sbjct 121 NVMDVLICGSMVSSLCFLGAIAVDRYVSIFYALRYHSIVTLPRAGRAIAAIWAGSVLSST 180


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Multiple alignment

Animal Genomics

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Animal Genomics

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Animal Genomics

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...CACGTGATGCGTACACTCGTGACGTGAACGGCGTC... DNA sequence

Animal Genomics

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Work flow

DNA preparation

Sequencing methods

Reading

DNA amplification

Animal Genomics

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ACGCTGCACGGCTG CGCGTACGTGCGTG ACGTGCGTGCGTGC ACGTGCGTGCACGC ACGTGGTGCAACAC ACGCTGCACGTGCA CGCGTACGTGCGTG ACGTGCGTGCGTGC ACGTGCGTGCACGC

CGCGTACGTGCGTG ACGTGCGTGCGTGC ACGTGCGTGCACGC ACGTGGTGCAACAC ACGCTGCACGTGCA CGCGTACGTGCGTG ACGTGCGTGCGTGC ACGTGCGTGCACGC ATGCCTCACGTGCT

ACGTGCGTGCGTGC ACGTGCGTGCACGC ACGTGGTGCAACAC ACGCTGCACGTGCA CGCGTACGTGCGTG ACGTGCGTGCGTGC ACGTGCGTGCACGCACGCTGCACGGCTG CGCGTACGTGCGTG

Generate whole-genome shotgun reads

Generate BAC shotgun reads

Animal Genomics

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(a) (b)

Animal Genomics

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Feb 2001 Feb 2001

Dec 2004

Dec 2005 Apr 2009

Jan 2010 Nov 2012

Animal Genomics

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Animal Genomics

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https://www.ncbi.nlm.nih.gov/genome/?term=chicken

Animal Genomics

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Animal Genomics

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Bos taurus (cow) is an agriculturally important animal; beef and milk production are the largest manufacturing industries in the United States. The cow is an important model organism for health research in obesity, female health, and infectious diseases.

Cow is also used in studies of endocrinology, physiology and reproductive techniques. The cow genome will be useful for comparative genomics, representing a non-primate, non-rodent eutherian genome. The B. taurus genome is estimated to be 3000 MB, organized in 29 pairs of autosomes and two sex chromosomes. Bovine chromosomes are acrocentric.

Animal Genomics

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https://www.ncbi.nlm.nih.gov/genome/?term=pig

Animal Genomics

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The giant panda, Ailuropoda melanoleuca, is a critically endangered mammal confined to portions of six isolated mountain ranges in south-central China. There are fewer than 3,000 giant pandas currently alive in the wild and fewer than two hundred in captivity. The giant panda is unusual for feeding almost exclusively on bamboo plants though being evolutionarily related to meat-eating carnivores such as bears and dogs.

(https://www.ncbi.nlm.nih.gov/genome/?term=panda)

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https://www.ncbi.nlm.nih.gov/genome/?term=sheep

Animal Genomics

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The ruminant goat is a member of the even-toed ungulates family Bovidae and is closely related to the sheep. Goats are one of the oldest domesticated species, and have been used for their milk, meat, hair, and skins over much of the world. Because of its long history of domestication, there are many different breeds which vary in size and weight, muscle mass, milk production, hair color and length, and horn shape and size . Goats are sexually dimorphic. Males have a beard, horns, a rank odor, and are generally larger than the females.

Animal Genomics

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The water buffalo Bubalus babalis is a domesticated animal that is agriculturally important as a source of meat and milk. The genome is organized in five pairs of metacentric chromosomes, 19 pairs of acrocentric chromosomes, and two sex chromosomes, X and Y.

626 6.40 Mb

677 8.72 Mb

738 8.71 Mb

Animal Genomics

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https://www.animalgenome.org/

Nature. 2010. 464:578-593

Animal Genomics

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626 6.40 Mb

677 8.72 Mb

738 8.71 Mb

Animal Genomics

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Genetic markers for improvement

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http://wallpapers.free-review.net/r?12

Phenotype

Genome Transcriptom

Proteome

DNA

(mRNA )

Bioinformatics

Conventional breeding M

olecular breeding Genetic markers for improvement

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Genetic marker

A

b

CTTCCTGAACGTCTATCAAGACCTGT

|||||||||||| |||||||||||||

CTTCCTGAACGTATATCAAGACCTGT

SNPs (single nucleotide polymorphisms)


a

B

CCGTAGTCGCGCGCGCGACTCTGTGG

||||||||||| |||||||||

CCGTAGTCGCG------ACTCTGTGG

Microsatellite (insertion/deletion)

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DNA markers for economic traits

A b

C

Genotype Markers

Phenotype

Marker b for gene A (closely linkage or linkage disequilibrium)

Marker C for gene A (unclosed linkage or linkage equilibrium)

Marker A for gene A (perfect marker located within gene sequence )

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DNA markers for economic traits

(a) Marker is gene responsible for traits and located within gene sequence

(b) Marker is not gene of interest, but it is closed or unclosed linked to gene

* * gene of interest

closed linked marker

test for variation

*

unclosed linked marker

test for variation

* gene of interest

test for variation

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(McPherron et al., 1997, Nature 387:83-90)P

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(McPherron et al., 1997, Nature 387:83-90)P AgBiotech - CMU


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Estrogen receptor gene (ESR) Follicle stimulating hormone beta (FSHB) Leukemia inhibitory factor gene (LIF)

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Candidate gene for litter size trait in pigs

Complement component factor B (BF) Erythropoietin receptor (EPOR) Retinol-binding protein 4 (RBP4)


Chromosome 1 (Ch1)

Ch2 Ch3 Ch4 Ch5

Ch 6 Ch7 Ch8 Ch9 Ch10

Quantitative trait loci (QTL) for economically important traits

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Genome wide association study (GWAS)

SNP chip

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Microarray

small-muscle Large-muscle


millions of DNA strands (oligos) build up one cell

labelled RNA fragments hybridize with DNA strands

Signal which is detected in the scanning process

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Differentially mRNA expression

Gene networks

Phenotype

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Differentially mRNA expression

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Gene networks

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Two Dimensional Electrophoresis (2-DE)

Peptide sequence (MALDI-TOF) mass spectrometry (Matrix assisted laser desorption /

ionization-time of flight)

Proteomics


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Proteomics is the study of the proteome or Total protein complement of a genome.

Protein solution

Isoelectric Focussing (IEF), first dimension

Vertical electrophoresis (2 nd dimension)

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Proteomic analysis (2 D – Electrophoresis)

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MW Proteomic analysis (2 D – Electrophoresis)


Protein solution

Isoelectric Focussing (IEF), first dimention

Vertical electrophoresis (2 nd dimension)

2 D - Electrophoresis

Low-performance High-performance

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Low –performance High –performance

N15 N15

N105 N105

N111 N111

Proteomic analysis AgBiotech - CMU


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Low- performance High- performance

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1 AAYRLVLVRH GESAWNLENR FCGWYDADLS PAGQQEARRG GEALRDAGYE

51 FDICFTSVQK RAIRTLWNVL DAIDQMWLPV VRTWRLNERH YGALTGLNKA

101 ETAAKHGEAQ VKIWRRSFDI PPPPMQSDHP FFSTISKDRR YADLTEDQLP

151 TCESLKDTIA RALPFWNEEI VPQIKEGKRV LIAAHGNSLR GIVKHLEGMS

201 EEAIMELNLP TGIPIVYELD KNLKPIKPMQ FLGDEETVRK AMEAVAAQGK

251 VKK

MALDI-TOF analysis

Proteomic analysis

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Form Proteomics to Genomics

1 AAYRLVLVRH GESAWNLENR FCGWYDADLS PAGQQEARRG GEALRDAGYE

51 FDICFTSVQK RAIRTLWNVL DAIDQMWLPV VRTWRLNERH YGALTGLNKA

101 ETAAKHGEAQ VKIWRRSFDI PPPPMQSDHP FFSTISKDRR YADLTEDQLP

151 TCESLKDTIA RALPFWNEEI VPQIKEGKRV LIAAHGNSLR GIVKHLEGMS

201 EEAIMELNLP TGIPIVYELD KNLKPIKPMQ FLGDEETVRK AMEAVAAQGK

251 VKK GGCGGTGCGGCGCGGGGCGGGTCAGTTGCGCGCGGCGGCGGCAGCGGGAGGAGCGCGAGAGCTGAGCGCCGGAGTCGCGATGGCCGCCTACCGC

CTCGTGCTCGTCCGCCACGGCGAGAGCGCCTGGAACCTGGAGAACCGCTTCTGCGGGTGGTACGACGCCGACCTCAGCCCCGCCGGCAGCAGGA

GGCGCGCCGCGGCGGAGAGGCCCTGCGAGACGCCGGCTATGAGTTCGATATCTGCTTCACGTCGGTGCAGAAGAGGGCCATCCGCACCCTCTGG

AACGTGCTGGACGCCATCGACCAGATGTGGCTGCCCGTGGTACGGACGTGGCGCCTCAACGAGCGGCACTACGGGGCCCTCACCGGCCTCAACA

AGGCCGAGACGGCCGCCAAGCACGGCGAGGCTCAGGTGAAGATCTGGAGGCGCTCCTTCGACATCCCCCCGCCGCCCATGCAGTCGGACCACCC

CTTCTTCAGCACCATCAGCAAGGATCGTCGCTACGCTGACCTGACGGAGGACCAGCTGCCTACATGTGAGAGCCTGAAGGACACCATCGCCCGG

GCCCTGCCCTTCTGGAATGAGGAAATAGTCCCGCAGATCAAAGAGGGCAAGAGAGTCCTTATCGCGGCCCATGGCAACAGCTTGCGGGGGATTG

TCAAGCATTTGGAAGGCATGTCAGAAGAGGCCATCATGGAACTGAACCTGCCCACCGGTATCCCAATCGTGTATGAATTGGACAAGAACCTGAA

GCCCATCAAGCCCATGCAGTTCCTAGGAGATGAGGAGACGGTGCGCAAGGCCATGGAGGCTGTCGCTGCTCAGGGCAAAGTCAAGAAATGAGGC

AGGCACGCAGACTAGCAAGTAGATGAGTCCCCTCCGTTCCCACCCCTCTGTGCACACCCCACAGCTGTAGGAACTTGGAGCTGCAGAGCTGGAG

CAGGGAGCAGCTCCCCAGGACTAGGTCCCTTCCCTCTGCCCCAGGCTCACCTCTGCAGGCAGCCTGGGAGCTGGGGGATGAGGGGCTGTGTGCA

CAGGGGTCTGGTGTGAAAGGGAGTCTTTGCTCTGCCAGGCAGCAACTGCAGCCCTATGTTCTGTGTTCCCAGCCCTTGGGGGGATGCTCTCGAG

CAATCTTATGCCATGCCTGGCTTCTCTTGCTTCCCTGGCAGCTCTTGTTAATGTTACTGTAGTTCTGTTGCTTGGTTTAGTTATTCCAGGAGCA

CAACGGGAGCAGTGGGACAGGGGCAGGAGGGGACAGTGACTAAGATGGGTGCCCAGTCCCCTCGTGGCTGTGTGGCACAAAGACTAGGATGCTT

AGACACCAAACGGGTTGTACTGTGGTGCATGCTCTTGTGTGTGGAACAGCACCCGGTAGTAGTACTTGCAAACCCTGACCCAGCTCTACCAGCC

AATGCTTTAACTTATTGTTTTTGGACAAAGGAAGGAGCGTGTTGTCTGGGAATTTGTATTTGCCATGTGCTTCCTACAGCCAAAGCCAACCCTA

GTCCTGCCTCACAGAGGTGGTGTAGATGCCATTGTGCCCCCACCCCACAAACGTGTACTTGGCACTGTCACTGTAGCCTCCCCTACTGCTGCTG

TAGTGAGCCTGGGCTTGCACTGGGTCAATCTAACTGTCTAGCGTTAGCAGTTGTCTTGTTCCAGTTCAATAAAAGATTAATATGCAAA

1

95

189

283

377

471

565

659

753

847

941

1035

1129

1223

1317

1411

1505

1599

Pyruvate kinase muscle 2 (PKM2)

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Genomics Technology

Transcriptom

Proteome

DNA

(mRNA )

Molecular breeding

Genome

Phenotype

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Biotechnology in Animal Reproduction

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Biotechnology in animal reproduction

Animal reproduction

Reproduction in mammals is achieved by each of two parents contributing genetic material to the young. Half of the characteristics of the young come from the father and half come from the mother.

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Reproductive technology in animals

Artificial insemination (AI)

In vitro production (IVP) of embryo

Cloning

Multiple ovulation and Embryo transfer (MOET)


Mating

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Mating


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Multiple ovulation – Embryo transfer

Superovulation (donor)

ET

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Recipient cows

offsprint

embryos


In vitro production of embryo

Oocyte collection (oocyte recovery)

In vitro maturation (IVM)

In vitro fertilization (IVF)

In vitro culture (IVC)

In vitro maturation, fertilization and culture

(IVMFC)

Embryo transfer

Embryo evaluation

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Ovum Pick Up (OPU)

pre-puberty puberty Non-pregnancy

pregnancy

killed animals

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Ovary of killed animals Ovary

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Oocyte collection with laparoscopy

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Ovum pick up (OPU) with ultrasound

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Ovum pick up (OPU)

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Embryo production

Immature Oocyte

IVM in media culture + 10µg of FSH (incubated for 22-24 hours at 39 oC in the humidified atmosphere containing 5% CO2)

IVF of matured Oocyte with capacitated sperm cells

IVC

(18 h)




Selection - egg cells

In vitro maturation of egg cells



In vitro fertilization (IVF)

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In vitro culture (IVC)

2-cell (22-32 hpi) 4-cell (32-48 hpi) 8-cell (48-65 hpi)

Blastocyst (140-170 hpi)

Matured oocytes

Morula (85-120 hpi) 16-cell (75-90 hpi)

Fertilization

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In vitro production of embryo

Oocyte collection


In vitro fertilization (IVF) In vitro culture

(IVC)

Embryo transfer

Embryo Evaluation

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Embryo evaluation

Good -quality Bad -quality



Embryo evaluation with sexing technique

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Development of animal reproductive technologies

Mating

A.I.

In vitro production embryo

Cloning

MOET

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Dolly was born on July 5, 1996 at the Roslin

Institute in Scotland. She was a clone, the first cloned sheep successfully born.

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The first genetically altered clone calves

(From: Herren, 2012. The Science of Agriculture. 4 edition, Delmer, New York)

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All eight of the calves were cloned from the same cow. (From: Herren, 2012. The Science of Agriculture A Biological Approach. 4 editon. Delmar, New York)

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Recombinant DNA Technology

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human

cloned human human protein AgBiotech - CMU



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- BST or growth hormone

- rBST is a genetically engineered version a of a hormone, which is found in the pituitary gland of cows and controls milk production.

- rBST can increase cows'milk production by as much as 20-30%.

Recombinant bovine somatotropin (rBST) hormone


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What is DNA vaccine?

- DNA vaccine is the vaccines which contain DNA that encodes for specific proteins (antigens) from a pathogen. - The DNA is injected into cells then used the DNA coding

to synthesize the protein - These proteins are recognized as foreign, when they processed the host cells and displayed on their surface and the immune will be responded.

DNA vaccine


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1. The RNA molecule is extracted from virus pathogen. RNA is converted to DNA.

2. The genetic sequence for virus is generated from the DNA.

3. The cDNA fragment will be amplified.

4.The cDNA fragment is inserted into plasmid.

5. The plasmids are grown in bacteria and purified by column chromatography 6. The purified DNA plasmids carrying the

virus gene make up to investigational vaccine.

Advantaes of DNA vaccine


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- Uses only the DNA from infectious organisms. - Avoid the risk of using directly pathogens. - Provide both humoral and cell mediated immunities. - Refrigeration is not required.

Transgenic Animals

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Transgenic Animals

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Reproductive technology Molecular technology

Superovulation donor

Recombination of gene construct Collection of embryos

DNA-microinjection

Embryo transfer

Transgenetic founder animals

Establishment of a transgenic population

Parturation of offspring DNA test

Expression analysis (RNA/protein)

Production of F1 generation

Breeding line

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Transgenic Animals

Micromanipulator

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Transgenic Animals

Microinjection

Pronucleus

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Transgenic Animals

Transgenic rat with human Growth hormone (hGH) (Palmiter et al. 1982)

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Transgenic Animals

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Transgenic Animals

Transgenic rat Wild type

Hakamata et.al. (2001) Biochem Biophys Res Commun. 286(4):779-85.

GFP expression pattern in GFP transgenic rat

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Transgenic Animals

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Transgenic Animals

Gong et al. (2003). Biochem. Biophys. Res. Commun. 308:58-63 AgBiotech - CMU

Transgenic Animals

Effeiciencies of gene transfer in animals

Animal born / injected embryo transferred

5-10 10-20 15-25 10-15 10-15 15 10-15

Transgenic animal /offspring (intergration freq.)

10-15 15-20 18 10-12 5-15 7 2-5

Pig Mouse Rat Rabbit Sheep Goat Cattle %

Transgenic / injected embryos transferred (efficiency)

0.5-1 2-3 4-5 1-2 1-2 1 0.2

(Brem, 1993; Brem and Mueller 1994; Wall, 1996)

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Transgenic Animals

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(a) (b) (c)

Transgenic Animals

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Transgenic Animals

Transgenic Animals

Protein Expressed Species Where Expressed Promoter

Reference

Lysozyme goat Bovine αs1-casein

(Maga et al. 2006)

Lysostaphin

cattle

Ovine β-lactoglobulin

(Wall et al. 2005)

Bovine β and κ casein

cattle

Bovine β-casein

(Brophy et al. 2003)

IGF-I

pig

Bovine α-lactalbumin

(Donovan et al. 2001)

α-lactalbumin

pig Bovine α-lactalbumin

(Bleck et al. 1998)

IGF-I

rabbits

Bovine αs1-casein

(Wolf et al. 1997)

Lactoferrin

cattle

Bovine αs1-casein

(Krimpenfort et al. 1991)

The target protein expression in milks of mamalian species

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Transgenic Animals

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Transgenic pig with phytase gene

to reduce phosphorus execrate from fecal animals to environment

Transgenic Animals

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Total phosphorus content (on a dry matter basis) of fecal matter from non-transgenic pigs ( ) and transgenic pigs ( ) of line WA fed different levels of soybean meal as the sole source of dietary phosphorus. DM, Dry matter content of feces.

Nature Biotechnology 19, 741–745 (1 August 2001) | doi:10.1038/90788

Introduction to Animal Biotechnology

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