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Molekular Genetika: Molekular TeknikEndang S. RahayuEndang S. Rahayu Fak.Teknologi Pertanian UGMAvery-MacLeod-McCarty experiment1944 Streptococcus pneumoniaEndang S. Rahayu Fak.Teknologi Pertanian UGMEndang S. Rahayu Fak.Teknologi Pertanian UGMMutasi, Recombinasi, Rekayasa Genetika• Mutasi, • Recombinasi– Transformasi – Transduksi – Konjugasi• Rekayasa GenetikaMutasiFigure 6.1Base-Pair SubstitutionsFigure 9.3Shifts in Reading FrameFigure 9.4RekombinasiGe
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Endang S. Rahayu
Fak.Teknologi Pertanian UGM
Molekular Genetika:
Molekular Teknik
Endang S. Rahayu
Endang S. Rahayu
Fak.Teknologi Pertanian UGM
Avery-MacLeod-McCarty experiment
1944 Streptococcus pneumonia
Endang S. Rahayu
Fak.Teknologi Pertanian UGM
Mutasi, Recombinasi, Rekayasa
Genetika
• Mutasi,
• Recombinasi
– Transformasi
– Transduksi
– Konjugasi
• Rekayasa Genetika
Mutasi
Figure 6.1
Figure 9.3
Base-Pair Substitutions
Figure 9.4
Shifts in Reading Frame
Rekombinasi
Figure 9.9
Genetic Recombination
Transformasi
Figure 9.10
Figure 9.14
Transformation
Transduksi
Figure 9.16
Generalized Transduction
Specialized Transduction
Figure 9.17
Konjugasi
Pla
sm
ids
Figure 9.23
Mechanism of Conjugation
Genetic Engineering Started in 1970s
Other names • Gene manipulation
• Recombinant DNA technology
• Recombinant DNA science
• Gene transfer
• Gene splicing
• Gene cloning
Genetic engineering • Technology involved in removing,
modifying, or adding genes to a DNA
molecule
• The transfer of genes (segment of
DNA) from one species to another.
• This is impossible in naturally
breeding
Gene manipulation
• The formation of new combinations of
heritable material by the insertion of
nucleic acid molecules, produced by
whatever means outside the cell, into any
virus, bacterial plasmid or other vector
system so as to allow their incorporation
into a host organism in which they do not
naturally occur but in which they are
capable of continued propagation
Endang S. Rahayu
Fak.Teknologi Pertanian UGM
Genetically Modified Organisms (GMOs)
Organisms that contain genetic
material that has been altered in
a manner that does not occur by
natural mutation, mating, or
recombination event
Endang S. Rahayu
Fak.Teknologi Pertanian UGM
Endang S. Rahayu
Fak.Teknologi Pertanian UGM
Endang S. Rahayu
Fak.Teknologi Pertanian UGM
Endang S. Rahayu
Fak.Teknologi Pertanian UGM
Products:
Recombinant Cultures
Endang S. Rahayu
Fak.Teknologi Pertanian UGM
Endang S. Rahayu
Fak.Teknologi Pertanian UGM
Topik
• Plasmids
• Restriction enzymes
• Gel electrophoresis
• Shouthern Blot
• PCR – Polymerase Chain Reactions
• Sequencing of the DNA
• Genetic Maps
Endang S. Rahayu
Fak.Teknologi Pertanian UGM
What is Plasmids ?
• Extrachromosomal DNA
• Having ori (origin of replication)
• Very small
• Carry genetic information
• Can be isolated from bacterial cells
• Important in a genetic engineering
Endang S. Rahayu
Fak.Teknologi Pertanian UGM
Natural role of a plasmid in bacteria?
• Plasmids usually contain one or two gene
that confer a selective advantage on the
bacterium e.g. antibiotic resistance gene.
• Every plasmid possess a replicator gene
can duplicated independently from the
chromosomal DNA.
Endang S. Rahayu
Fak.Teknologi Pertanian UGM
Antibiotic resistance genens and
replicator gene
Endang S. Rahayu
Fak.Teknologi Pertanian UGM
Plasmid pBR322
Endang S. Rahayu
Fak.Teknologi Pertanian UGM
Endang S. Rahayu
Fak.Teknologi Pertanian UGM
Endang S. Rahayu
Fak.Teknologi Pertanian UGM
Endang S. Rahayu
Fak.Teknologi Pertanian UGM
Endang S. Rahayu
Fak.Teknologi Pertanian UGM
Restriction Enzymes
Endang S. Rahayu
Fak.Teknologi Pertanian UGM
DNA double helixes are cut
at the axis of symmetry:
Endang S. Rahayu
Fak.Teknologi Pertanian UGM
DNA double helixes are cut
: sticky end
Endang S. Rahayu
Fak.Teknologi Pertanian UGM
Endang S. Rahayu
Fak.Teknologi Pertanian UGM
Endang S. Rahayu
Fak.Teknologi Pertanian UGM
Endang S. Rahayu
Fak.Teknologi Pertanian UGM
Endang S. Rahayu
Fak.Teknologi Pertanian UGM
EcoR1 Recognition Site
Figure 6.13
Endang S. Rahayu
Fak.Teknologi Pertanian UGM
Endang S. Rahayu
Fak.Teknologi Pertanian UGM
Electrophoresis
Endang S. Rahayu
Fak.Teknologi Pertanian UGM
Endang S. Rahayu
Fak.Teknologi Pertanian UGM
Endang S. Rahayu
Fak.Teknologi Pertanian UGM
Endang S. Rahayu
Fak.Teknologi Pertanian UGM
Endang S. Rahayu
Fak.Teknologi Pertanian UGM
Endang S. Rahayu
Fak.Teknologi Pertanian UGM
Endang S. Rahayu
Fak.Teknologi Pertanian UGM
Southern Blot
Endang S. Rahayu
Fak.Teknologi Pertanian UGM
Endang S. Rahayu
Fak.Teknologi Pertanian UGM
Endang S. Rahayu
Fak.Teknologi Pertanian UGM
Endang S. Rahayu
Fak.Teknologi Pertanian UGM
Polymerase Chain Reaction
Endang S. Rahayu
Fak.Teknologi Pertanian UGM
Endang S. Rahayu
Fak.Teknologi Pertanian UGM
Sequencing the DNA
Endang S. Rahayu
Fak.Teknologi Pertanian UGM
Endang S. Rahayu
Fak.Teknologi Pertanian UGM
Endang S. Rahayu
Fak.Teknologi Pertanian UGM
Genetic maps of E. coli
Endang S. Rahayu
Fak.Teknologi Pertanian UGM