Mikrobiologi Bioteknologi

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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

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