CLONING TECHNOLOGY Matakuliah Genetika (BIO612206) Jurusan Biologi Fakultas MIPA Universitas Lampung [email protected] staff.unila.ac.id/priyambodo
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CLONING
TECHNOLOGY
Matakuliah Genetika (BIO612206)
Jurusan Biologi Fakultas MIPA Universitas Lampung
[email protected] staff.unila.ac.id/priyambodo
(1) Rekombinasi DNA (Kloning DNA)
(2) Kloning reproduktif
(3) Kloning terapeutik
Types of Cloning Technology
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Setiap gen dapat diisolasi dan sekuens DNA dari gen tersebut dapat ditentukan urutannya
Kloning DNA:
Memperbanyak sekuens DNA tertentu untuk tujuan tertentu
Kloning DNA
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• The names of restriction enzymes are generally derived from the names of the bacteria from which they were originally isolated.
• Thus, HindIII and EcoRI are derived from Hemophilus influenzae and E. coli, respectively.
Restriction enzymes
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Cutting DNA
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• Many restriction enzymes, such as HindIII, cut DNA at sequences that are palindromic, producing complementary sticky ends. The small black arrows designate the enzyme’s cleavage sites.
• For bacteria vectors are based on DNA molecules that move between cells in nature - bacterial viruses (bacteriophages/phages) and plasmids.
• Mammalian vectors usually derive from mammalian viruses.
• In higher plants, the favored system is the infectious agent of crown-gall tumors (Ti plasmids from Agrobacterium tumefaciens) or of adventitious roots (Ri plasmids from A. rhizogenes).
Vektor kloning
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• Plasmid • Bacteriophage • Cosmid
A cloning vector consisting of the phage lambda cos site (the sequence that is cut to produce the cohesive, single-stranded extensions located at the ends of the linear DNA molecules of certain phages) inserted into a plasmid. Such vectors can be packaged into lambda phage or maintained as plasmids.
• Yeast Artificial Chromosome (YAC) A cloning vector which contains sequences from a yeast chromosome required for DNA replication and segregration. Often used for cloning very large fragments of DNA.
• Bacterial Artificial Chromosome (BAC) Low copy number plasmid vectors that allow stable cloning very large DNA fragments (often 100 Kb or more).
Cloning vector
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Vector Insert size (kb)
Plasmid <10 kb
Bacteriophage 9-20 kb
Cosmids 33-47 kb
BACs 75-125 kb
YACS 100-1000 kb
Cloning vector
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How is DNA cloned?
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In 1981, a new series of plasmids were developed that permitted the identification of the foreign DNA containing cells in a single screening step.
• Is a technology used to generate an animal that has the same nuclear DNA as another currently or previously existing animal.
• Dolly was created by reproductive cloning technology, in a process called "somatic cell nuclear transfer" (SCNT)
Reproductive cloning
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• Also called "embryo cloning"
• Is the production of human embryos for use in research.
• The goal of this process is not to create cloned human beings, but rather to harvest stem cells that can be used to study human development and to treat disease.
Therapeutic cloning
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– A normal allele could be inserted by a viral vector into some bone marrow cells removed from the patient.
– If the procedure succeeds, the returned modified cells will multiply throughout the patient’s life and express the normal gene, providing missing proteins
Stem cell technology
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Embryonic Reproductive Cloning Laws Worldwide
Embryonic Research Cloning Laws Worldwide
Referensi
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Albert B, Johnson A, Lewis J, Raff M, Roberts K, and Walter P (2002) Molecular Biology of the Cell. Garland Science, taylor & Francis Group
Brooker RJ(1999) Genetics: Analysis and Principles. Addison-Wesley Febriyanto, Y.H. (2006) Somatic cell nuclear transfer (SCNT). Kuliah Perdana Program Studi
Bioteknologi UGM 2006. Nelson DL and Cox MM (2000) Lehninger Principles of Biochemistry. Worth Publisher Marzuki, S (2006) Bio-Engineering dan Umat manusia Kuliah Perdana, Sekolah Pascasarjana,
Universitas Gajah Mada Yogyakarta, 6 September 2006 Forum on Science Ethics and Policy (FOSEP – at UW) http://www.fosep.org/ A Small Dose of Toxicology www.asmalldoseof.org The bioscience-bioethics friendship Co-operative http://www.bioscience-bioethics.org/portal.htm Richard Deem,vidence for God from Science (www.GodAndScience.org)
Ahmad Aulia Jusuf. 2009, Bagian Histologi, Fakultas Kedokteran Universitas Indonesia
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