Kuliah Pengantar Bioteknologi Kehutanan Pokok Bahasan 1 Pendahuluan - Pengertian dan Sejarah - Prospek dan Tantangan Pokok Bahasan 2 Genetika Molekuler Tanaman - Materi Genetik - Regulasi dan Ekspresi Gen - Perubahan Materi Genetik Pokok Bahasan 3 Rekayasan Gen - Kloning-transformasi - Metode-transformasi Pokok Bahasan 4 Sistem Silvikultur Jaringan - Mikropropagasi dan Kultur Sel - Kultur Haploid dan Variasi Somaklonal Pokok Bahasan 5 Penerapan Bioteknologi - Identifikasi dan Klasifikasi Molekuler (Isoenzym, RFLP, RAPD, sidik jari) - Perlindungan terhadap Hama & Penyakit - Organisme Pemacu Pertumbuhan Tanaman - Organisme Transgenik
Bioteknologi : Teknologi yang menggunakan sistem hayati (proses-proses biologi) untuk mendapatkan barang dan jasa yang berguna bagi kesejahteraan manusia Pengertian dan Sejarah
Sejarah Minuman alkohol anggur (pengawetan daging) Khamir roti Keju Yogurt Susu masam kecap 1857 – Pasteur menemukan bahwa fermentasi merupakan proses yang dilakukan oleh organisme hidup 1920 – fermentasi aseton, etanol, butanol, gliserin PD II – Penicillium natatum Antibiotik Vitamin, steroid, enzim
Amerika Serikat mengijinkan digunakannya sel tubuh manusia untuk kegiatan penelitian mengenai terapi gen.1990 Metode polymerase chain reaction (PCR) pertama kali dipublikasikan1988 Badan paten Amerika Serikat mengumkan bahwa tikus yang direkayasa secara genetis peka terhadap kanker1988 Ti plasmid yang telah direkayasa digunakan untuk transformasi pada tanaman1983 Pertama kalinya vaksin hewan diproduksi menggunakan teknologi DNA rekombinan di Eropa1982 Pertama kalinya kit diagnostik yang dibuat dari antibodi monoklonal digunakan secara luas di Amerika Serikat1981 Pertama kalinya alat otomatis untuk sintesis DNA dijual secara komersial1981 Badan pengadilan Amerika Serikat menetapkan peraturan dalam kasus Diamond VS Chakrabarty yang melakukan manipulasi mikroorganisme yang dipatenkan. 1980 Perusahaan Genentech memproduksi insulin manusia pada sel bakteri Escherichia coli1978 Pengembangan teknik untuk mengetahui sekuen DNA1976 Pertama kali dikeluarkan pedoman untuk penelitian dalam bidang DNA rekombinan1976 Kohler dan Milstein mendeskripsikan produksi antibodi monoklonal1975 Boyer dan Cohen mengembangkan teknologi DNA rekombinan1973 Khorana dan rekan-rekannya dapat mensintesis keseluruhan gen yang mengkode tRNA1972 Isolasi pertama kali enzim endonuklease restriksi1970 Keseluruhan kode genetik dapat diketahui1961-1966 Watson dan Crick menemukan struktur DNA1953 Avery, MacLeod dan McCarty membuktikan bahwa DNA merupakan materi genetik.1944 Antibiotik penisilin diproduksi besar-besaran dalam skala industri1943 Karl Ereky pertama kali menyatakan istilah bioteknologi1917 PeristiwaTahun
Bioteknologi tradisional : Seleksi bahan, mikrobia yaang digunakan dan modifikasi lingkungan untuk memperoleh produk optimal. Misal : pembuatan tempe, tape, roti, pengomposan sampah Bioteknologi modern : Memanfaatkan ketrampilan manusia dalam melakukan manipulasi makhluk hidup agar dapat digunakan untuk menghasilkan suatu barang yang diinginkan. Misal rekayasa genetik Pengertian & Sejarah
Bioteknologi merupakan penerapan prinsip ilmiah dan rekayasa pengolahan bahan oleh agen biologi untuk menyediakan barang dan jasa Rekayasa genetik : Teknik untuk menghasilkan molekul DNA yang berisi gen baru yang diinginkan atau kombinasi gen-gen baru atau dapat dikatakan sebagai manipulasi organisme Pengertian & Sejarah
Hasil-hasil rekayasa genetika tanaman yang digambarkan dalam National Enquier, c. 1981.
Prospek & Tantangan Pertimbangan Pemanfaatan Hasil Bioteknologi 1) Tinjauan ekologik a)GMO mengalahkan spesies alami, menggantikan spesias alami, mengurangi keanekargaman hayati. b)GMO berubah prilaku dibandingkan dengan aslinya. c)Gen yang disisipkan dapat pindah ke kerabat dekat spesies GMO, termasuk karakteristik gen yang dimaksud. Penambahan gen karakter baru, kemampuan baru, kemampuan hayati baru.
Kesetabilan ‘gen asing’ atau dapat berubah dan diekspresikan berbeda oleh organisme yang tertular gen asing – mutasi Penarikan ‘gen asing’ jika masuk dalam organisme non target Terganggunya ‘rantai makanan' Terganggunya organisme non target
2) Tinjauan kesehatan a)Gangguan kesehatan karena produk gen asing (dalam waktu panjang) b)Bahaya atau tidaknya gen-gen penyerta 1) Tinjauan sosial dan budaya a)Etika, moral b)Agama 2) Perdagangan global
The History of Biotechnology http://www.i-s-b.org/wissen/timeline/englisch/
1859 Charles Darwin publishes his book (On the Origin of Species by Means of Natural Selection). According to this book evolution is life´s motor. The interplay of mutation and selection endows living beings with optimized traits in order to survive. These principles are also valid for the so-called 'chemical evolution' of biomolecules and are being used in laboratories for in vitro optimizing of wanted qualities in moleculesOn the Origin of Species by Means of Natural Selection
1865 Gregor Mendel finds that independent 'factors' are responsible for the heredity of traits from one generation to the next according to a set of (Mendelian) priciples.
1869 Friedrich Miescher discovers an acidic substance in the nucleus of cells which he names 'nuclein'. By elemental analysis he finds 14 % nitrogen, 3 % phosphorus, and 2 % sulfur (from proteins). Since the substance cannot be cleaved by the proteolytic enzyme pepsin Miescher concludes that this substance is not a protein.
1879 Walther Flemming observes the separation of chromosomes during mitosis but fails to fully understand its meaning. He counts 24 pairs of chromosomes, a number which will be corrected to 23 later in 1956 by the Indonesian scientist Joe- Han Tijo.1956
1900 William Bateson introduces 'genetics' as a scientific discipline Hugo de Vries, Erich Tschermak von Seysenegg and Carl Correns independently rediscover the Mendelian principles Hugo de Vries defines the meaning of 'mutations'
1902 Walter Sutton observes in grasshoppers' cells that chromosomes carry the Mendelian 'factors of heredity', i.e. the genetic information.
1909 Wilhelm Johansen coins the terms 'gene', 'genotype', and 'phenotype' referring to Mendel's 'factors'. Archibald Garrod confirms the hereditary nature of four metabolic diseases. He publishes "Inborn Errors of Metabolism"hereditary nature of four metabolic diseases
1910 Thomas Hunt Morgan's studies in the biology of Drosophila melanogaster confirm that certain traits are inherited sex- specifically. He also proves that some phenotypes result from several genes located on different chromosomes.
1927 Herman Muller finds that energetic radiation causes defects in the chromosomes, i.e. mutations.
1928 Fred Griffith demonstrates that material from killed bacteria of the pathogenic strain Streptococcus pneumoniae S ('smooth') is taken up from living ones of the non-pathogenic strain Streptococcus pneumoniae R ('rough') which are subsequently 'transformed' into the pathogenic S-strain. Pneumococcus pneumoniae
1944 Oswald Theodore Avery, Colin McLeod, and Maclyn McCarty find by careful analysis of Griffith' experiments that desoxynucleic acid (DNA) is the carrier of the 'transforming principle'.
1945 Erwin Schrödinger proposes in his famous book 'What is life?' that genes must be 'aperiodic crystals' consisting of a succession of a small number of isomeric elements whose precise sequence constitutes the heredity code. Although these ideas do nothing to identify the responsible molecular structures, they attract many newcomers to the field.
1950 Erwin Chargaff observes the 1:1 ratio of the nucleic bases adenine/thymine and guanine/cytosine. This will be a decisive hint for Watson & Crick to the structure of the DNA molecule. Watson & Crick
1952 Alfred Day Hershey's and Martha Chase's studies in bacteriophages prove that only the nucleic acids carry the genetic information; proteins are definitively excluded from playing this role. Joshua Lederberg finds plasmids in bacteria, small rings of extrachromosomal DNA which can be replicated autonomously.
1953 James D. Watson and Francis H. C. Crick elucidate the structure of the DNA molecule. It consists of two strands which are bound by hydrogen bonds bewtween opposite basepairs of adenine-thymine and cytosine-guanine. Their model is based on the results of many colleages, namely Rosalind Franklin who discovered the helical structure and the outward position of the phosphate-sugar backbone. Rosalind Franklin
1955 Frederick Sanger finds a method to sequence proteins. With this he determines the sequence of the amino acids of insulin.
1956.. Arthur Kornberg reports the first in vitro synthesis of a DNA molecule. George Emile Palade locates protein biosynthesis to the ribosomes, the 'factories of the cell'.
1956 Joe Han Tijo and Albert Levan correct the number of the chromosome pairs in human cells to 23. Since the days of Walther Flemming 24 was believed to be the true number.Walther Flemming Chromosomen einer menschlichen Zelle protein chains are built at the ribosomes
1959 T. Akiba, T. Koyama, Y. Isshiki, S. Kimua, T. Fukushima, T. Watanabe and T. Fukusawa describe the transfer of multiple resistance to antibiotics by bacterial plasmids. Plasmids turn out to be efficient vectors for trafficking genetic information between bacteria of different strains. bacterial plasmid (arrow)
1960 Francois Jacob and Jaques Monod recognize the function of the messenger RNA (mRNA). They develop the operon model of gene regulation in procaryotes.operon model
1961 Sydney Brenner and Francis Crick claim that every amino acid corresponds to a triplett of nucleotides, called 'codons'.Francis Crick Marshall Nirenberg and Heinrich Mathaei can prove that the codon UUU in mRNA codes for the amino acid phenyl alanine Sydney Brenner Marshall Nirenberg
1962 John Gurdon claims to have reproduced frogs from the epithelium cells of Xenopus laevis. J. B. S. Haldane coins 'cloning' when describing these debated experiments. Xenopus laevis South African clawed frogs John Gurdon
1966 Mainly due to the work of Har Gobind Khorana the 'Genetic Code' is completely known.
1968 Werner Arber discovers enzymes, so-called nucleases, which digest DNA double strands from the ends.
1969 Jonathan Beckwith is the first to isolate a complete gene, in this case a gene from the bacterial sugar metabolism.
1970 Hamilton Smith discovers an enzyme (HindII) which specifically cleaves DNA double strands Har Gobind Khorana synthesizes a complete gene (that of an alanine t-RNA) in vitro. David Baltimore and Howard Temin discover the viral enzyme reverse transcriptase. It translates RNA sequences to DNA sequences and thus tumbles the age old 'Central Dogma of Genetics' which defined the direction of the flow of the genetic information to start from DNA via RNA to the proteins.
1971 Paul Berg, Peter Loban, and Dale Kaiser find enzymes which attach short single strands to the blunt ends of double stranded DNA. The single strands consist of only one sort of nucleotides. Complementary single strands (sticky ends) allow the coupling of ds DNA fragments; the remaining gaps can be closed by the use of ligases.
1972 Janet Mertz and Ron Davis paste DNA fragments of different origin resulting from specific enzymatic restriction. They close the gaps using ligases and thus produce recombinant DNA.
1973 Stanley Cohen and Herbert Boyer paste enzymatically restricted DNA fragments into plasmids which have been cleaved by restriction enzymes. The resulting recombinant plasmids can serve as vectors for the transfer of foreign DNA into bacteria. These experiments are generally regarded as the beginning of the era of genetic engineering and modern biotechnology.
1975 The first conference on safety issues of the new technology is held at Asilomar in California. Georges Köhler and Cesar Milstein develop the 'Hybridoma Technology' for the production of monoclonal antibodies. An antibody producing plasma cell (derived from a B lymphocyte) is fused with a tumor cell resulting in an immortal hybridoma cell which (as well as its daughters) continues to produce one kind of antibodies.
1976 Herbert BoyerHerbert Boyer and Robert Swanson found Genentech the first biotech company.
1977 Walter Gilbert and Frederick Sanger independently develop two different methods for DNA sequencing.Frederick Sanger
1978 Walter Gilbert discovers that genes of eucaryotic organisms are composed of coding and non-coding parts. The non-coding sequences, named introns, are cut from the mRNA before translation leaving the coding exons as the source of information for protein synthesis.
1979 Thomas Cech and Sydney Altman discover the autocatalytic activity of some RNA molecules to cleave themselves at well defined positions. They coin 'ribozyme' for this type of RNA.
1980 Jozef Schell and Marc van Montagu transfer foreign DNA into plant cells by employing T- plasmids of Agrobacterium tumefaciens as vectors.
1981 The first patent for a genetically modified organism is granted to Ananda Chakrabarty of General Electric. It covers a P. aeruginosa strain which is equipped with genes of certain enzymes in order to metabolize crude oil.
1982 The U.S. Food and Drug Agency (FDA) approves recombinant insulin for marketing.
1984 Alec Jeffreys develops 'genetic fingerprinting' allowing the comparison of minute variations, the so-called restriction length fragment polymorphisms (RLFPs), in the genomes of two organsims.
1985 Karry Mullis develops the polymerase chain reaction (PCR). This powerful tool allows to copy and accumulate extremely low amounts of DNA from various sources until a level sufficient for analysis is reached.
1986 Neal First develops protocols for clonig cattle from bovine embryos by separating cells at early embryonic stages. These experiments resemble John Gurdons earlier attempts at the cloning of frogs. The first cloning of an adult animal will be reported in 1997 by Ian Wilmut who succeeds in cloning a sheep, Dolly.John GurdonsIan Wilmut
1988 The first patent for a transgenic mammal is granted to Harvard's Philip Leder and Timothy Stewart. It refers to a mouse which is susceptible to tumors and serves as model organism for studies in cancer.
1990 French Anderson successfully tries the first somatic gene therapy of a human. Patient Ashanti DeSilva is cured from an deficiency in the enzyme ADA, which causes severe immunodeficiency. German 'Genetic Engineering Act' passes parliament Official kick-off of the Human Genome Project (HGP)
1993 The German Genetic Engineering Act of 1990 is revised, i.e. many restrictions of the 1990 version are eased.Genetic Engineering Act1990 The Biotechnology Industry Organization BIO is created by merging two smaller trade associationsBiotechnology Industry Organization BIO
1994 Genetically modified tomatoes are on sale in the U.S. Cans containing puree from transgenic tomatoes are on the shelves of british supermarkets.
1995 The Institute for Genomic Research (TIGR) publishes the first complete sequence of the genome of a free-living organism, the bacterium Haemophilus influenzae Germany launches the BioRegio contest. 17 regions compete in presenting the best blueprint for the development of commercial biotechnology to an international jury. The three winning regions will share the price of DM 150m.BioRegio
1996 The complete sequencing of the baker's yeast' genome is accomplished by close collaboration of laboratories in Europe, Japan, and the U.S. The Saccharomyces cervisiae genome ist the first eucaryotic genome completely sequenced. Patrick Browne of Stanford University presents the first 'gene chip' containing 6116 different gene specific sequences of the baker's yeast genome. In the U.S. transgenic plants grow on more than 1.9 million hectares
1997 Ian Wilmut (right) visits Dolly (center) the first mammal cloned from an adult cell.
1998 Two research teams report embryonic stem cells differentiate to specialized tissue cells. The picture shows an early developmental stage, the blastocyst. The first genome of a multicellular organism (Caenorhabditis elegans) is sequenced. Craig Mello und Andrew Fire find that small double-stranded RNA molecules can selectively block gene expression in Caenorhabditis elegans, a phenomenon later referred to as RNA interference (RNAi)
1999 Pluripotent stem cells from tissues can be reprogrammed to other cell types. The sequence of chromosome 22 is published. The U.S. biotech company Celera starts its own human genome sequencing project
2000 There are 1300 biotech companies on each side of the Atlantic. About 400 are listed at stock exchanges. The genome of Drosophila melanogaster is sequenced. The fruit fly was introduced by Thomas H. Morgan into genetics research. The Human Genome Organization and Celera present the first working draft of the human genome. The picture shows Celera's CEO J. Craig Venter (left) and Francis Collins, the speaker of the HGP.
2001… Less genes than expected The international Human Genome Project (HGP) and biotech company Celera Genomics publish their versions of the sequence of the human genome. The Celera team estimates the number of genes to range from 26,000 to 39,000, HGP scientist' estimate is between 30,000 and 40,000. Rice genome Syngenta (Basel, Switzerland), in collaboration with Myriad Genetics (Salt Lake City, UT), announced the completed sequencing of the genome of Oryza sativa japonica
2001 RNA interference Thomas Tuschl is the first to demonstrate that RNA interference (RNAi) is also working with mammalian cells.
2002 Genomes of the malaria agent and its carrier sequenced Researchers of the Institute for Genomic Research (TIGR), the Sanger Institute (UK), and Stanford University publish the genomic sequence of Plasmodium falciparum and Plasmodium yoelii. The protozoa P. falciparum causes malaria. The genome of its carrier Anopheles gambiae is also sequenced and published.
2003… Genome of SARS virus sequenced Within three weeks after its discovery scientists of the Michael Smith Genome Sciences Centre in British Columbia (Vancouver, Canada) succeeded in sequencing the genome of the virus causing severe acute respiratory syndrome (SARS). The RNA virus remotely resembles corona virus species and contains roughly 30,000 genes.
2003 Germ cells from stem cells Karin Hübner and Hans Schöler of the University of Pennnsylvania succeeded in generating oocytes from murine stem cells which were able to grow into further embryonic stages. The experiment demonstrates the potential of pluripotent stem cells to acquire totipotency via a germ line cell stage
2004… 99.999% accuracy Three years after the publication of the first draft a far more precise version of the sequence of the human genome is published. The number of identified genes decreases again compared with previous estimations. 19,599 have been confirmed to encode proteins, 2188 are suspected to encode for proteins. 1183 genes have been generated through duplication of genes, 33 genes appear to have been degenerated into disfunctional pseudogenes.
2004… Fatherless mouse Tomohiro Kono of the Tokyo University of Agriculture presented the first mouse which was obtained from oocytes only. His team combined the genomes of oocytes from adult and newborn mice and eventually one of 460 experiments resulted in a living animal, being 14 month old at the time of publication. Critical for success was the suppression of a gene which normally controls the imprinting of chromosomal DNA from mother and father.
2004 Mice from cancer cells Rudolph Jaenisch of MIT and Lynda Chin of the Dana Farber Cancer Institute generated cloned mice from cancer cells by stuffing the nuclei of melanoma cells into de- nucleated mouse oocytes. The resulting blastocysts yielded stem cells which were suitable for transplantation into normal blastocysts which eventually developed into living animals. Some genetic markers of the melanoma cells were found again in the animal, but the epigentic patterns had gone lost - showing that epigenetic modifcations are both reversible and late stage in the transformation of normal cells into melanoma cells.