poulry genetics, new techniques of molecular genetics

1. Advances in molecular genetics of poultry Presented By : Alok Bharti MVSc. [ AGB ]

5. Early Vision on Molecular Biology

9. Molecular genetics tools Molecular Genetics Cloning Technology DNA Sequencing Technology Transgenesis Technology PCR Technologies Marker Assisted Selection

12. Why do we care about variations? underlie phenotypic differences allow tracking human/animal history (ancient and modern) cause inherited diseases

21. RAPD: Randomly amplified polymorphic DNA Size sorted

23. RAPD Analysis B

28. AFLPs

31. VNTR Mini satellite

36. What is a SNP?

39. SNP Types

42. Polymerase Chain Reaction

45. Standard Polymerase Chain Reaction

46. PROCEDURE DENATURATION ANNEALING DNA SYNTHESIS

52. Denaturation Annealing Extention

67. How DNA Sequence Is Determined? Polyacrylamide Gel Electrophoresis T A G C T A C G DNA fragments having a difference of one nucleotide can be separated on gel electrophoresis But these bands can’t tell us the identity of the terminal nucleotides If those band with the same terminal nucleotide can be grouped, then it is possible to read the whole sequence Juang RH (2004) BCbasics ATC 32 P AT 32 P A 32 P ATCG 32 P ATCGA 32 P ATCGAT 32 P ATCGATC 32 P ATCGATCG 32 P ATCGATCGA 32 P ATCGATCGAT 32 P

68. How to Obtain DNA Fragments AT ATCGAT Specific Reaction to G A Terminated Biosynthetic method Chemical method Template or Non-radioactive (invisible) Producing various fragments

69. Phosphodiester bond 3’ 5’ di deoxy nuceotide Terminated dd NTP Sanger’s Method: How Terminated Normal Linking Can not react Juang RH (2004) BCbasics P R P R P R P R P R P R OH 5’ 3’ 1 A 1 2 3 4 5 6 A PO 4 2- H 3’ 5’ 2 H

71. GENE CLONING

72. Gene Transferred by Plasmid Plasmid gets out and into the host cell Resistant Strain New Resistance Strain Non-resistant Strain Plasmid Enzyme Hydrolyzing Antibiotics Juang RH (2004) BCbasics Drug Resistant Gene mRNA

73. Target Genes Carried by Plasmid 1 plasmid 1 cell Recombinant Plasmid Transformation Target Gene Recombination Restriction Enzyme Restriction Enzyme Chromosomal DNA Target Genes DNA Recombination Transformation Host Cells

74. Amplification and Screening of Target Gene 1 1 cell line, 1 colony X100 X1,000 Plasmid Duplication Bacteria Duplication Plating Pick the colony containing target gene =100,000

82. Fig 22.13 The production of transgenic animals by microinjection of DNA into fertilized eggs. © 2003 John Wiley and Sons Publishers

87. Why Express rProtein in Milk Easy to purify - few other proteins in milk Doesn’t harm transgenic animal- no change to physiology rProtein is authentically modified post-translationally Large quantities Renewable source

88. Sheep and Pigs PIGS PST porcine somatotropin (growth hormone) adverse effects- kidney, stomach, heart, sterility human Hemoglobin to replace whole blood transfusions SHEEP Increase wool production keratin promoter growth factor

89. Organ Transplant Pr Problem: Rejection Antibodies from Host bind to Donor Organ Elicits Inflammatory Response Transplanted Organ Lost Solution: Transgene in Donor for Complement-Inhibiting Protein 111

90. Birds and Fish Birds traditional methods can not be used because of avian embryogenesis differences no ES cells found ALV resistant chickens transgene - defective ALV genome makes viral RNA and protein but blocks assembly of viral particles Fish aquaculture transgene - growth hormone

105. Every body fishing for New Ideas

106. How much we differ from Ape DNA answers ?

107. April 25th celebrates the DNA Model DNA continues to the Temple of Science

108. Madam, are you fiddling with GENES ?

109. Be A GENE Genius Become Famous