BIOLOGI MOLEKULER Pengenalan Biologi Molekuler Materi Genetik

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BIOLOGI MOLEKULER Pengenalan Biologi Molekuler Materi Genetik Struktur dan Sifat Asam Nukleat Replikasi DNA, Mutasi dan Repair DNA III. Ekspresi Gena Transkripsi, Translasi, dan Regulasi IV. Pertumbuhan Sel Eukariot Sel Signaling dan Siklus Sel V. Transformasi Genetik Rekombinasi, Transposon dan Retrovirus

III. DAFTAR PUSTAKA Lodish, H., Berk, A., Zipursky, A.L., Matsudaira, P., Baltimore, D., Darnell, J., 2000, Molecular cell Biology, 4th ed., Freeman and Company, New York. Alberts,B., Bray, D., Lewis, J., Raff, M., Robert, K., Watson, JD., 1994, Molecular Biology of the Cell, 3th ed.,Garland Publishing, Inc, New York. Devlin T.M., 2002, Biochemistry with Clinical Correlations, 5th ed., wiley-Liss, New York. Berg, J.M., Tymoczko, J.L., Stryer.,2002, Biochemistry, 5th ed.,W.H. Freeman and Company, New York. Freifelder, D.,1987, Molecular Biology, 2th ed., Jones and Bartlett Publisher, Boston. Glick, B.R. and Pasternak, J.J., 1998, Molecular Biotechnology, Principles and Applications of Recombinant DNA, 2nd. Ed. ASM Press, Washington DC. Brown, T.A., 2001, Gene Cloning and DNA Analysis, An Introduction, 4th ed., Blackwell Science, UK

mempelajari struktur & fungsi Biologi Molekuler mempelajari struktur & fungsi DNA, RNA , protein 46 Human 2.9x109 bp (Berg JM et al, 2002)

FARMASI BIOLOGI MOLEKULER PROTEIN OBAT REKAYASA GENETIK PENEMUAN OBAT TANAMAN TRANGENIK VAKSIN PROTEIN OBAT ALAT DETEKSI PENYAKIT KIMIA MEDISINAL FARMAKOLOGI BIOKIMIA MIKROBIOLOGI BIOLOGI MOLEKULER FARMASI REKAYASA GENETIK TEKNOLOGI GENA FARMASETIK

PERBEDAAN PROKARIOT & EUKARIOT Organisme Bakteria dan sianobakteria Fungi, hewan, manusia Ukuran sel 1 - 10m 5 - 100m Organel Beberapa / tidak ada Inti, mitokondria, kloroplast GENOM Sirkuler dalam sitoplasma, Satu molekul kromosom Linier panjang, dipak rapi dgn protein histon dalam inti. Manusia mempunyai 46 cromosom RNA dan protein Disintesis dalam sitoplasma RNA : dalam inti, Protein: dalam sitoplasma Organisasi Unisel Multisel

mRNA Translation Proteins Decay DNA mRNA IFs, EF, RF RNAses Regulatory factors Proteins Decay Turnover Interactions Localization Modification

(PO4/dPO4, glycosylation) Pre-mRNA Capping Polyadenylation Splicing mRNA eIFs, eEF, eRF Storage mRNA Translation RNAses Regulatory factors Proteins Decay Turnover Interactions Localization Modification (PO4/dPO4, glycosylation)

II. MATERI GENETIK Mahluk hidup tersusun dari  SEL yang dapat membelah dan menurunkan INFORMASI GENETIK nya Dibawa oleh DNA : rantai polimer panjang yang merupakan rangkaian dari jutaan nukleotida Fragmen DNA yang mengkode protein, suatu unit keturunan  Gena Molekul DNA yang terdiri dari beberapa gena di paking membentuk CHROMOSOM Total informasi genetik yang tersimpan dalam kromosom  GENOM

What Is Cloning? A clone is a genetically identical copy of a living organism, such as the famous sheep ‘Dolly’ Similar to an identical twin human clones are never absolutely identical to the original; for example, so-called ‘identical’ human twins, while clones, are still two different people who possess their own separate souls

The Making of Dolly Empty DNA DNA Sheep Egg Mother’s Egg Cloning depends on DNA Step 1: Take the Nuclei out of a sheep egg Step 2: Transfer nuclei from the Mother Empty DNA DNA The mother is to be cloned since her DNA was used Sheep Egg Mother’s Egg

She was cloned using a cell taken from a healthy six-year-old sheep Hello Dolly July 5, 1996, Dolly was born at the Roslin Institute, Edinburgh, Scotland She was cloned using a cell taken from a healthy six-year-old sheep

Cloning and the Cell Nucleus Its all about DNA In cloning we are interested in the Nucleus (DNA) Its all about DNA

Cloning Depends on DNA Nucleus Chromosome Each cell contains DNA. DNA is packaged into compact units called chromosomes. Basically a chromosome is one long chain of genetic material. It is the microscopic, rod-shaped, threadlike part of the cell that carries hereditary information in the form of genes.  Every cell has chromosomes and all individuals in the same species have the same number of chromosomes.  Within an individual, every cell has the same number of chromosomes and generally come in pairs (except in sex cells). People with Down Syndrome have 3 copies of chromosome 21. telomeres, these molecular chains have often been compared to the blank leaders on film and recording tape. Indeed, telomeres seem to perform a similar function in aligning the DNA molecule during the replication process. Protecting the vital DNA molecule from being copied out of synch, these telomeres provide a kind of buffer zone where asynchronous replication errors (that are inevitable) will not result in any of the vital DNA sequences being lost. As a cell gets older, it is under attack by oxides and other so-called free-radical chemicals in the body and environment. We survive as living beings because our cells have the ability to duplicate themselves before being killed by these natural causes. Each time our cells duplicate themselves, a small portion of the DNA molecule is lost and not copied. The loss is usually to the telomere and so the effect is usually insignificant. Scientists recently noted that the length of these telomere chains were shorter as we grew older. Eventually, the telomeres become so shortened that the losses in replication begin to effect the vital DNA molecule sequence and prevent the cell from being able to duplicate itself. This is why we age. Chromosome

Good-Bye Dolly It’s all about DNA At age 3 Dolly showed signs of premature aging. Dolly died at 6 years old from progressive lung disease (symptom of old age) Dolly’s DNA was already 6 years old when she was born - WHY? “Dolly’s birth was one of the biggest news stories of the nineties. Scientists made extravagant promises about medical and technological advances. Well, Dolly’s embarrassingly premature death received little attention, precisely because it exposes the horrors of cloning.” Dolly was diagnosed with arthritis, normally found in old sheep Telomeres are the ends of our chromosomes. Made of many repeats of the same DNA sequence, they act like shoelace caps: they protect the gene-containing body of the chromosome from being worn down. Breakdowns in telomere maintenance are implicated in ageing and cancer. Telomeres sit on the ends of chromosomes to protect them from damage. When chromosomes are replicated during cell division, a stretch of the telomere is left unreplicated—making the telomere a bit shorter with each division. After some 50 to 100 divisions, the telomeres become so short that the cell can no longer divide—a phenomenon scientists call senescence—that is, the state of being old. It’s all about DNA

Telomeres TTAGGG Telomeres Gene 1 Gene 2 DNA Chromosome TELOMERES in human beings are a repeating sequence of three of these nucleotide units: Thymine (T), Adenine (A), and Guanine (G), in the sequence: Important in replication - they act like a buffer zone Each time a normal cell divides its telomeres are cut shorter Cells with critically short telomeres become unable to divide further, and eventually malfunction and die Basically a chromosome is one long chain of genetic material. It is the microscopic, rod-shaped, threadlike part of the cell that carries hereditary information in the form of genes.  Every cell has chromosomes and all individuals in the same species have the same number of chromosomes.  Within an individual, every cell has the same number of chromosomes and generally come in pairs (except in sex cells).   Humans, for instance, have 23 pairs, or 46 chromosomes. A gene can be defined as a region of DNA that controls a hereditary characteristic. It usually corresponds to a sequence used in the production of a specific protein or RNA. Genes can be as short as 1000 base pairs or as long as several hundred thousand base pairs. It can even be carried by more than one chromosome. The estimate for the number of genes in humans has decreased as our knowledge has increased. As of 2001, humans are thought to have between 30,000 and 40,000 genes. telomere is a repeating DNA sequence (TTAGGG) at the end of the body's chromosomes. The telomere can reach a length of 15,000 base pairs. Telomeres function by preventing chromosomes from losing base pair sequences at their ends. They also stop chromosomes from fusing to each other. However, each time a cell divides, some of the telomere is lost (usually 25-200 base pairs per division). When the telomere becomes too short, the chromosome reaches a "critical length" and can no longer replicate. This means that a cell becomes "old" and dies by a process called apoptosis. Telomere activity is controlled by two mechanisms: erosion and addition. Erosion, as mentioned, occurs each time a cell divides. Addition is determined by the activity of telomerase. apoptosis: The process by which a cell dies at a natural, "pre-programmed" time Chromosome

Telomere Length Declines 1,500 3,000 8,000 Age (years) Telomere length in base pairs (human white blood cells) 35 Telomere length declines in dividing cells as we age. In human blood cells, the length of telomeres ranges from 8,000 base pairs at birth to 3,000 base pairs as people age and as low as 1,500 in elderly people. (An entire chromosome has about 150 million base pairs.) Each time a cell divides, an average person loses 30 to 200 base pairs from the ends of that cell’s telomeres. Cells normally can divide only about 50 to 70 times, with telomeres getting progressively shorter until the cells become senescent, die or sustain genetic damage that can cause cancer. Telomeres do not shorten with age in tissues such as heart muscle in which cells do not continually divide. 65

More on Dolly and Cloning Dolly’s experimenters used 277 cloned embryos to produce one sheep, meaning 276 failed. Question How many failures will it take to produce a human A telomere is a repeating DNA sequence (TTAGGG) at the end of the body's chromosomes. The telomere can reach a length of 15,000 base pairs. Telomeres function by preventing chromosomes from losing base pair sequences at their ends. They also stop chromosomes from fusing to each other. However, each time a cell divides, some of the telomere is lost (usually 25-200 base pairs per division). When the telomere becomes too short, the chromosome reaches a "critical length" and can no longer replicate. This means that a cell becomes "old" and dies by a process called apoptosis. Telomere activity is controlled by two mechanisms: erosion and addition. Erosion, as mentioned, occurs each time a cell divides. Addition is determined by the activity of telomerase. apoptosis: The process by which a cell dies at a natural, "pre-programmed" time ?

Other Cloned Animals July 1998 cloned calves Cloned mouse Cloned mule

Future Experiments ButterCat CatCow

Future Experiments SparrowBoxer TigerOwl

JUMLAH BASA VS KOMPLEKS MAKHLUK

Chromosomes 16 6 24

Chromosomes 78 40 46

Human Genome Genome inti Genome mitokondria 23 pasang kromosome  2x (4x109 bp)  2 m DNA/sel Genome mitokondria

Some organisms are diploid - that is, they have paired homologous chromosomes in their somatic cells, and thus contain two copies of each gene. An organism in which both copies of the gene are identical - that is, have the same allele - is said to be homozygous for that gene. An organism which has two different alleles of the gene is said to be heterozygous

Masing2 single linier DNA Human 4 x 109  22 autosom yang berbeda 2 sex kromosom Masing2 single linier DNA E. Coli 4.7 x 106  1 Chromosom 40 – 50 loop superkoil

Kromosom tanpa histon

DNA CHROMOSOME

KROMOSOM CENTROMER pengikatan mitotic spindle saat proses mitosis region dari chromosom yang diperlukan untuk pengikatan mitotic spindle saat proses mitosis TELOMER fragmen di ujung chromosom linier replikasi secara khusus untuk mencegah perpendekan chromosom ORIGIN REPLIKASI tempat awal replikasi berlangsung

Introduction The Central Dogma of Molecular Biology Cell DNA mRNA Transcription Reverse tanscription Polypeptide (protein) Translation Ribosome

KOMPOSISI GENA FUNGSIONAL PROMOTER OPERATOR ACTIVATOR BINDING SITE STRUKTURAL GEN : mRNA TERMINATOR

Teknik yang sering digunakan Isolasi DNA & RNA Elektroforesis Hibridisasi PCR Sekuensing DNA