Farmasi UGM BIOLOGI SEL MOLEKULAR farmasibugm 2012gmail com

Farmasi UGM BIOLOGI SEL & MOLEKULAR farmasibugm 2012@gmail. com 28/03/2014 MODIFIED by Prof. Sismindari 1

Kuliah ke Dosen Pokok Bahasan Sub Pokok Bahasan I. Prof. Sismindari Pengenalan Biologi molekuler Peranan Biologi Molekuler dalam Farmasi, - - Contoh perkembangan dalam Biologi Molekular : kloning domba Dolly dan terapi sel induk materi genetik : Genom, kromosom, gena, DNA, RNA, protein. Perbedaan genom prokariot dan mamalia, lokasicentromer, telomere, dan origin replikasi, human genom project- Dogma sentral ekspresi gen: replikasi, transkripsi, translasi- Organisasi dari sel Struktur dan fungsi organela sel Arsitektur sel: Cytoskeleton & Cell Junction materi genetik : Genom, kromosom, gena, DNA, RNA, protein. Struktur DNA (model Watson dan Crick) dan RNA, basepairing DNA, Sifat DNA dan RNA Heterochromatin, Karyotyping Chromosome Pertumbuhan, Perkembangan dan Diferensiasi Sel - Penurunan sifat materi genetic, Pertumbuhan Sel Pembelahan Sel, meiosis dan mitosis (metaphase, anaphase, interfase) Hukum Mendel Diferensiasi Sel, Komunikasi Antar Sel Replikasi DNA (Sintesis DNA) -Reaksi 1) II. 2) + 3) Dr. Puji A. III. 4) +5) Prof Sismindari 28/03/2014 polimerisasi, Proses replikasi DNA: konservatif dan semi-konservatif, inisiasi, elongasi replikasi, - Garpu replikasi, fragmen Okazaki, peran telomere, peran enzim MODIFIED by Prof. Sismindari 2 topoisomerase, mekanisme proofreading, mutasi karena triple repeat.

Kuliah ke Pokok Bahasan Sub Pokok Bahasan Dosen III. 5) Prof Sismindari Mutasi dan Repair DNA - Definisi, mutan, mutasi, mutagen; jenis-jenis mutasi, jenis-jenis mutan, jenis-jenis mutagen (kimia, fisika dan biologi), mutasi pada level gen, pada level organisme. Stabilitas genetik dan kerusakan DNA, macamkerusakan DNA, proses umum reparasi DNA, penyakit yang disebabkan oleh adanya mutasi III. 6) Prof Djadi Polimorfism SNP kaitannya dengan keragaman genetik masing individu: Farmakogenomik aplikasinya pada metabolisme INH dan warfarin Polimorfisme mikrosatelit : Sidik jari DNA aplikasinya pada hubungan kekerabatan dan identifikasi individu dengan PCR Hibridisasi, macam dan prinsip kerjanya (Southern, Northen) 7) Mekanisme infeksi virus dan Transformasi genetik - Macam-macam virus (virus DNA, virus RNA), genom virus, Life cycle HIV, Virus influenza, EBV, dan retrovirus Jenis transposon: transposon, viral retrotransposon dan non-viral retrotransposon. - Mekanisme transposisi MODIFIED by Prof. Sismindari Penyakit yang disebablkan oleh 3 transposon 28/03/2014

Dosen Pokok Bahasan Sub Pokok Bahasan IV. Prof Kuswandi 8) Transkripsi pada prokariot : RNA polimerase: inisiasi (TATA Box, promoter), elongasi, terminasi. Fenomena transkripsi pada prokariot : polisistronik, polisom, repressor, aktivator dan induser Fenomena transkripsi eukariot: intron, ekson, capping, splicing dan poli-A, monosistronik Obat-obat yang bekerja pada penghambatan proses transkripsi Ekspresi Gen: Transkripsi, prokariot, eukariot Regulasi: Sistem operon, Lac repressor dan Catabolic activator protein (CAP), triptofan operon 9) Ekspresi Gen: Ribosom, kodon antikodon , t. RNA, aminoasyl t. RNA sintetase Proses : inisiasi, elongasi, dan terminasi Perbedaan proses translasi pada prokariot dan eukariot. Pasca translasi : modifikasi asam amino. Obat-obat dengan target penghambatan proses translasi prokariot (antibiotik). Teknik deteksi ekspresi gen V. Dr Riris 28/03/2014 10) Sistem regulasi ekspresi gen factor transkripsi, domain dan motif, contoh factor MODIFIED by Prof. Sismindari 4 transkripsi, dan peran factor transkripsi. Signal transduksi sederhana (hormone steroid), Translasi

Dosen Pokok Sub Pokok Bahasan Sistem regulasi melalui second messenger G-protein signaling : G-protein, jalur c. AMP, jalur protein kinase C (PKC) Aplikasi regulasi pengendalian ekspresi gen pada proses pengobatan 12) Prof. Edy M Cell-cycle dan regulasinya Overview : tahapan pembelahan sel dan daur sel (G 1 S-G 2 -M-phases), Kontrol daur sel pada G 1 -phase (peran cyclin-cdk dan p. Rb), pengaturan sistem checkpoint pada daur sel Apoptosis (kematian sel terprogram) : jenis kematian sel, karakteristik morfologi apoptosis, jalur instrinsik dan ekstrinsik, peran pro dan anti apoptosis (bcl 2 famili) Peran mitokondria. Teknik deteksi apoptosis VII. 13) 14) Karsinogene sis Pengertian dasar kanker, tahap-tahap karsinogenesis, golongan agen karsinogenik (kimia, fisika, dan biologi), pengenalan marker molecular kanker (mis. Ras dan p 53) dan perannya pada proses karsinogenesis, pengembangan agen kemoprevensi 11) Dr. Riris 28/03/2014 MODIFIED by Prof. Sismindari 5

III. DAFTAR PUSTAKA Lodish, H. , Berk, A. , Zipursky, A. L. , Matsudaira, P. , Baltimore, D. , Darnell, J. , 2000, Molecular cell Biology, 4 th ed. , Freeman and Company, New York. Alberts, B. , Bray, D. , Lewis, J. , Raff, M. , Robert, K. , Watson, JD. , 1994, Molecular Biology of the Cell, 3 th ed. , Garland Publishing, Inc, New York. Devlin T. M. , 2002, Biochemistry with Clinical Correlations, 5 th ed. , wiley-Liss, New York. Berg, J. M. , Tymoczko, J. L. , Stryer. , 2002, Biochemistry, 5 th ed. , W. H. Freeman and Company, New York. Glick, B. R. and Pasternak, J. J. , 1998, Molecular Biotechnology, Principles and Applications of Recombinant DNA, 2 nd. Ed. ASM Press, Washington DC. Sismindari, 2012, Seri Biologi Molekuler: Replikasi DNA, Pustaka Pelajar, Yogyakarta 28/03/2014 MODIFIED by Prof. Sismindari 6

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• Peranan Biologi Molekuler dalam Farmasi. • Materi genetik : Genom, kromosom, gena, DNA, RNA, protein. • Perbedaan genom prokariot dan mamalia, lokasi centromer, telomere, dan origin replikasi, human genom project • Dogma sentral ekspresi gen : replikasi, transkripsi, translasi • Contoh perkembangan dalam Biologi Molekular : kloning domba Dolly dan terapi sel induk 28/03/2014 MODIFIED by Prof. Sismindari 8

How to do well in Mol Biol • Study every day • Don’t just look at notes, rewrite, reorganize, redraw pictures from memory • When read book, take notes • Ask questions • Come to office by appointment 28/03/2014 MODIFIED by Prof. Sismindari 9

Human 2. 9 x 109 bp 46 28/03/2014 MODIFIED by Prof. Sismindari 10 (Berg JM et al, 2002)

Farmasi UGM Molecular Biology The field overlaps with other areas of biology, particularly genetics and biochemistry Molecular biology concerns itself with understanding the interactions between the various systems of a cell, including the interrelationship of DNA, RNA and protein synthesis and learning how these interactions are regulated. Function Biochemistry Proteins Genetics Genes Molecular Biology 11 Schematic relationship between biochemistry, genetics and molecular biology MODIFIED by Prof. Sismindari introduction%20 to%20 molecular%20 and%20 cell%20 biology. ppt 28/03/2014

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Mengapa mempelajari 28/03/2014 MODIFIED by Prof. Sismindari 13

MANFAAT BIOLOGI MOLEKULER GENOME MUTASI DNA PADA GENOME REPLIKASI DNA TRANSKRIPSI TRANSLASI 1. SIFAT & FUNGSI BERUBAH 2. SIFAT & FUNGSI TETAP • TERJADINYA PENYAKIT • PERUBAHAN RESPON OBAT MODIFIED by Prof. Sismindari PCR SEKUENSING • PROTEIN, RESEPTOR • TARGET OBAT • PROTEIN OBAT TARGET TERAPI 28/03/2014 14

MANFAAT BIOLOGI MOLEKULER REGULASI PROKARIOT REGULASI EUKARIOT REGULASI SECOND MESSENGER REGULASI SIKLUS SEL MODIFIED by Prof. Sismindari - RESPON BAKTERI THDP LINGKUNGAN - MEMPRODUKSI PROT OBT DLM BAKTERI - RESPON HUMAN THDP LINGKUNGAN - PROSES METABOLISME - SAAT TERJADI INFEKSI PATOGEN • BAGAIMANA SIGNAL DARI LUAR DITERUSKAN KE DLM SEL • SIGNAL DI DITERUSKAN MELALUI RESEPTOR • BAGAIMANA PEMBELAHAN SEL NORMAL DIATUR AGAR TDK TERJADI KANKER 28/03/2014 15

KETERKAITAN DENGAN ILMU LAIN 28/03/2014 MODIFIED by Prof. Sismindari 16

Farmasi UGM What is Life made of: • All living things are made of Cells – Nucleus = library – Kromosom = bookshelves – Genes = books Almost every cell in an organism contains the same libraries and the same sets of books. Nucleus Books represent all the information (DNA) that every cell in the body needs can grow and carry out its various functions. DEVIDED & PASSED THE GENETIC INFORMATION in to the doughter cells 28/03/2014 • All Life depends on 3 critical molecules From DNA, to RNA, to Proteins MODIFIED by Prof. Sismindari 17

PERBEDAAN PROKARIOT & EUKARIOT 28/03/2014 MODIFIED by Prof. Sismindari 18

PERBEDAAN PROKARIOT & EUKARIOT 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 28/03/2014 MODIFIED by Prof. Sismindari 19

Eukariot Prokariot Splising 28/03/2014 MODIFIED by Prof. Sismindari 20 (Alberts B, et al, 1994)

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Farmasi UGM DNA, RNA, and the Flow of Information Functional Proteins Replication Post translation modification Transcription 28/03/2014 MODIFIED by Prof. Sismindari Translation 22

PEMBELAHAN SEL 28/03/2014 MODIFIED by Prof. Sismindari 23

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FENOTIP 28/03/2014 MODIFIED by Prof. Sismindari 25

DNA CHROMOSOME Kromosom tanpa histon 28/03/2014 MODIFIED by Prof. Sismindari 26

KROMOSOM CENTROMER 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 28/03/2014 MODIFIED by Prof. Sismindari 27

Telomere Length Declines 8, 000 Telomere length in base pairs (human white 3, 000 blood cells) 1, 500 28/03/2014 MODIFIED by Prof. Sismindari 0 35 65 Age (years) 28

Chromosomes 16 78 28/03/2014 24 6 46 40 MODIFIED by Prof. Sismindari 29

Human 4 x 109 22 autosom yang berbeda 2 sex kromosom Masing 2 single linier DNA diploid 2 copies ech chromosome 2 copies each gene 1 X chrom in male E. Coli 4. 7 x 106 1 Chromosom 40 – 50 loop superkoil 28/03/2014 MODIFIED by Prof. Sismindari 30

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Aplikasi biomol 1 28/03/2014 MODIFIED by Prof. Sismindari 32

You can tell people apart by their fingerprints… Because everyone’s fingerprints are different! 28/03/2014 MODIFIED by Prof. Sismindari 33

DNA is like a fingerprint because everyone’s is a little different! How does the police look at DNA to figure out who committed a crime? 34 MODIFIED by Prof. Sismindari 28/03/2014

Miss Ellis 28/03/2014 Sara This is what it really looks like!!! MODIFIED by Prof. Sismindari 35

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Medicine and the New Genetics Gene Testing � Pharmacogenomics � Gene Therapy Anticipated Benefits: • improved diagnosis of disease • earlier detection of genetic predispositions to disease • rational drug design • gene therapy and control systems for drugs • personalized, custom drugs 28/03/2014 MODIFIED by Prof. Sismindari 38 U. S. Department of Energy Genome Programs, Genomics and Its Impact on Science and Society, 2003

Aplikasi 2 POLIMORFISME 28/03/2014 MODIFIED by Prof. Sismindari 39

Pharmacogenomics Same symptoms, Same findings, Same disease? Different patients Same drug Same dose Why does drug response vary? Genetic Differences SNP G Different Effects Ethnicity Age Weight Pregnancy Possible Reasons: Individual variation By chance… Genetic factors A Disease Drug interactions. . 28/03/2014 MODIFIED by Prof. Sismindari 40

Single Nucleotide Polymorphisms (SNPs) A key to human variability DNA sequence variation at a single nucleotide that may alter the function of the encoded protein * Functional protein Functional but altered protein Polymorphisms are common and contribute to common diseases and influence our response to medications 28/03/2014 MODIFIED by Prof. Sismindari 41

Phase II enzymes known to have polymorphism î NAT 2(N-acethyl Transferase): Isoniazid, hydralazine, î GST: D-Penicillamine î TPMT: Azathioprine, 6 -MP î Pseudocholinesterase: Succinyl choline î UGT 1 A 1: Irinotecan 42 MODIFIED by Prof. Sismindari 28/03/2014

"Here's my sequence. . . " The New Yorker 28/03/2014 MODIFIED by Prof. Sismindari 43

APLIKASI BIOMOL UNTUK KLONING 28/03/2014 MODIFIED by Prof. Sismindari 44

The Making of Dolly Cloning depends on DNA Step 1: Take the Nuclei out of a sheep egg Step 2: Transfer nuclei from the Mother DNA Empty DNA Sheep Egg 28/03/2014 Mother’s Egg MODIFIED by Prof. Sismindari 45

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 28/03/2014 MODIFIED by Prof. Sismindari 46

Good-Bye Dolly At age 3 Dolly showed signs of premature aging. u 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? u It’s all about DNA 28/03/2014 MODIFIED by Prof. Sismindari 47

Other Cloned Animals July 1998 cloned calves 28/03/2014 Cloned mouse MODIFIED by Prof. Sismindari Cloned mule 48

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KOMPOSISI GENA FUNGSIONAL PROMOTER OPERATOR ACTIVATOR BINDING SITE STRUKTURAL GEN : m. RNA TERMINATOR 28/03/2014 MODIFIED by Prof. Sismindari 50

Introduction The Central Dogma of Molecular Biology Cell Transcription Translation Reverse tanscription DNA m. RNA Ribosome Polypeptide (protein) 28/03/2014 MODIFIED by Prof. Sismindari 51

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ASISITENSI ASISTEN BIOLOGI SEL & MOLEKULER 28 Februari 2014 28/03/2014 MODIFIED by Prof. Sismindari 56

TATA TERTIB LABORATORIUM • Mahasiswa Datang 15 menit sebelum praktikum dimulai • Jas laboratorium dan name tag dipakai sebelum memasuki laboratorium • Mahasiswa tidak diperkenankan pakai SANDAL dan KAOS OBLONG • Mahasiswa terlambat 15 menit tanpa alasan yang jelas tidak diperkenankan mengikuti praktikum • Mahasiswa diwajibkan untuk MENGIKUTI SEMUA MATA PRAKTIKUM • Tidak disediakan waktu khusus untuk inhal. • Mahasiswa yang berhalangan hadir bisa mengikuti PRAKTIKUM GOLONGAN LAIN untuk acara yang sama • Mahasiswa tidak diperkenankan MAKAN, MINUM, MEROKOK selama praktikum • Laporan praktikum dibuat BER-KELOMPOK dan dikumpulkan saat mengikuti praktikum berikutnya • 28/03/2014 MODIFIED by Prof. Sismindari 57

Percobaan • Percobaan 1 : Isolasi DNA MAMALIA • Percobaan 2 A : KUANTIFIKASI DNA HASIL ISOLASI DENGAN SPEKTROFOTOMETRI • Percobaan 2 B : AMPLIFIKASI DNA HUMAN DENGAN PCR UNTUK MELIHAT POLIMORFISME • Percobaan 3 : ANALISIS DNA DGN ELEKTROFORESIS GEL AGAROSE • Percobaan 4: PENGAMATAN SEL DENGAN MIKROSKOP • Percobaan 5: PENGENALAN BIOINFORMATIKA BLAST • Percobaan 6: SIMULASI DETEKSI EKSPRESI PROTEIN DENGAN TEKNIK WESTERN BLOT 28/03/2014 MODIFIED by Prof. Sismindari 58

Penilaian • • Test : 30 Kerja : 10 Laporan : 20 Responsi: 40 BOBOT 1 • NILAI UJIAN (UTS+UAS) BOBOT 3 • NILAI AKHIR {(3 XNILAI UJIAN)+NILAI PRAKTIKUM}/4 28/03/2014 MODIFIED by Prof. Sismindari 59

SISTEM EVALUASI • NILAI AKHIR – – – 28/03/2014 A B C D E > 75 65 – 74 55 – 64 35 – 54 <34 MODIFIED by Prof. Sismindari 60