CLONING TECHNOLOGY

Cell. Garland Science, taylor & Francis Group Brooker RJ(1999) Genetics: Analysis and Principles. Addison-Wesley Febriyanto, Y.H. (2006) Somatic cell ...

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Matakuliah Genetika (BIO612206) Jurusan Biologi Fakultas MIPA Universitas Lampung

CLONING TECHNOLOGY

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Types of Cloning Technology (1) Rekombinasi DNA (Kloning DNA) (2) Kloning reproduktif (3) Kloning terapeutik

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Kloning DNA 

Setiap gen dapat diisolasi dan sekuens DNA dari gen tersebut dapat ditentukan urutannya



Kloning DNA: 

Memperbanyak sekuens DNA tertentu untuk tujuan tertentu

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Restriction enzymes • 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.

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Restriction enzymes

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Cutting DNA • 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.

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Vektor kloning • 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). staff.unila.ac.id/priyambodo

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Cloning vector • 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).

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Cloning vector Vector

Insert size (kb)

Plasmid

<10 kb

Bacteriophage

9-20 kb

Cosmids

33-47 kb

BACs

75-125 kb

YACS

100-1000 kb

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Recombinant DNA technology

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How is DNA cloned? In 1981, a new series of plasmids were developed that permitted the identification of the foreign DNA containing cells in a single screening step.

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Microinjection

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Reproductive cloning • 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)

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Therapeutic cloning • 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.

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Stem cell technology

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Stem cell technology – 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 staff.unila.ac.id/priyambodo

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Stem cell technology

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Embryonic Reproductive Cloning Laws Worldwide

Embryonic Research Cloning Laws Worldwide

For further info...

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Referensi 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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