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PERAN GEN DALAM PERKEMBANGAN • Para peneliti mengklon hewan melalui transplantasi inti – Inti sel telur digantikan oleh inti sel somatik dewasa • Sejauh ini, kloning pada manusia belum berhasil hingga menghasilkan embrio kurang dari 6 sel – Perkembangan embrio ditentukan oleh ekspresi gen • Dalam reproduksi eukariota secara kloning, embrio diimplantasikan di induknya • Untuk terapi, kloning dihasilkan dari stem sel embrionik – Stem cells can help patients with damaged tissues Donor cell Nucleus from donor cell Remove nucleus from egg cell Add somatic cell from adult donor Implant blastocyst in surrogate mother Clone of donor is born (REPRODUCTIVE cloning) Remove embryonic stem cells from blastocyst and grow in culture Induce stem cells to form specialized cells for THERAPEUTIC use Grow in culture to produce an early embryo (blastocyst) GENE REGULATION IN PROKARYOTES Interaksi protein dan DNA pada gen prokariotik: on-off tergantung pada respon perubahan lingkungan • Aliran informasi genetik dari gen ke protein disebut dengan ekspresi gen ▫ Penjelasan awalnya menggunakan pengaturan gene dari bakteri E. coli • Pada prokariot, gen berperan sebagai enzim, pengaturannya secara bersama menjadi unit pengaturan yang disebut operon • Pengaturan pengikatan protein yang mengontrol urutan DNA dan on-off kerja operon diatur oleh perubahan lingkungan • Operon lac akan menghasilkan enzim untuk mengurai laktose jika ada laktose OPERON Regulatory gene Promoter Operator Lactose-utilization genes DNA mRNA RNA polymerase cannot attach to promoter Active repressor Protein OPERON TURNED OFF (lactose absent) DNA RNA polymerase bound to promoter mRNA Protein Lactose Inactive repressor Enzymes for lactose utilization OPERON TURNED ON (lactose inactivates repressor) • Dua tipe repressor yang dikontrol oleh operon Promoter Operator Genes DNA Active repressor Active repressor Tryptophan Inactive repressor Inactive repressor Lactose lac OPERON trp OPERON Diferensiasi sel dan kloning eukariota • Pada eukariota, sel menjadi spesifik seperti zigot berkembang menjadi organisme ▫ Tipe sel yang berbeda menyebabkan perbedaan macam protein penyusunnya ▫ Perbedaan kombinasi gen penyusun protein: aktif setiap tipe sel • An overview of cellular respiration Electrons carried via NADH and FADH2 Electrons carried via NADH Citric acid cycle Glycolsis Pyruvate Glucose Cytosol Mitochondrion ATP Figure 9.6 Oxidative phosphorylation: electron transport and chemiosmosis Substrate-level phosphorylation ATP Substrate-level phosphorylation ATP Oxidative phosphorylation 1 When blood glucose level rises, a gland called the pancreas secretes insulin, a hormone, into the blood. 2 Insulin enhances the transport of glucose into body cells and stimulates the liver and muscle cells to store glucose as glycogen. As a result, blood glucose level drops. STIMULUS: Blood glucose level rises after eating. Homeostasis: 90 mg glucose/ 100 mL blood 4 Glucagon promotes the breakdown of glycogen in the liver and the release of glucose into the blood, increasing blood glucose level. STIMULUS: Blood glucose level drops below set point. 3 When blood glucose level drops, the pancreas secretes the hormone glucagon, which opposes the effect of insulin. • Awal eksperimen: transplantasi inti sel hewan (katak) – Kloning larva katak: inti sel somatik berpotensi genetik dan berkembang menjadi embrio selanjutnya larva katak Tadpole (frog larva) Frog egg cell Nucleus UV Intestinal cell Nucleus Transplantation of nucleus Nucleus destroyed Tadpole Eight-cell embryo • The first mammalian clone, a sheep named Dolly, was produced in 1997 – Dolly provided further evidence for the developmental potential of cell nuclei Reproductive cloning of nonhuman mammals has applications in basic research, agriculture, and medicine • Scientists clone farm animals with specific sets of desirable traits • Piglet clones might someday provide a source of organs for human transplant stem sel dapat diperbanyak dan dapat digunakan untuk terapi setelah mengalami diferensiasi sel • Stem sel dewasa dapat di kultur dan mengalami diferensiasi • Diferensiasi sel dipengaruhi oleh perubahan media kultur • Differentiation of embryonic stem cells in culture Liver cells Cultured embryonic stem cells Nerve cells Heart muscle cells Different culture conditions Different types of differentiated cells GENE REGULATION IN EUKARYOTES DNA packing in eukaryotic chromosomes helps regulate gene expression • A chromosome contains a DNA double helix wound around clusters of histone proteins • DNA packing tends to block gene expression DNA double helix (2-nm diameter) Histones “Beads on a string” Nucleosome (10-nm diameter) Tight helical fiber (30-nm diameter) Supercoil (200-nm diameter) 700 nm Metaphase chromosome In female mammals, one X chromosome is inactive in each cell • An extreme example of DNA packing in interphase cells is X chromosome inactivation EARLY EMBRYO TWO CELL POPULATIONS IN ADULT Cell division and X chromosome inactivation X chromosomes Allele for orange fur Allele for black fur Active X Inactive X Inactive X Active X Orange fur Black fur Chromosome DNA unpacking Other changes to DNA GENE GENE TRANSCRIPTION Exon RNA transcript Intron Addition of cap and tail Splicing Tail Cap mRNA in nucleus NUCLEUS Flow through nuclear envelope mRNA in cytoplasm CYTOPLASM Breakdown of mRNA Translation Broken-down mRNA Polypeptide Cleavage/modification/ activation ACTIVE PROTEIN Breakdown of protein Broken-down protein The Control of Gene Expression • Mutant fruit flies show the relationship between gene expression and development – Some mutants have legs where antennae should be Eye Antenna Head of a normal fruit fly Leg Head of a developmental mutant • Development of head-tail polarity in fruit fly EGG CELL WITHIN OVARIAN FOLLICLE Egg cell Egg protein signaling follicle cells 1 Follicle cells 2 Gene expression in follicle cells Follicle cell protein signaling egg cell Localization of “head” mRNA 3 “Head” mRNA ZYGOTE FERTILIZATION AND MITOSIS EMBRYO Translation of “head” mRNA Gradient of regulatory protein 4 Gene expression 5 Gradient of certain other proteins Gene expression Body segments 6 EMBRYO Body segments 6 LARVA Gene expression ADULT FLY 7 Head end Tail end SIGNALING CELL A signal-transduction pathway that turns on a gene 1 2 TARGET CELL (1) The signaling cell secretes the signal molecule (2) The signal molecule binds to a receptor protein in the target cell’s plasma membrane Signal molecule Receptor protein Plasma membrane SIGNALING CELL Signal molecule 1 (3) Binding activates the first relay protein, which then activates the next relay protein, etc. 2 Plasma membrane 3 TARGET CELL Relay proteins Transcription factor (activated) (4) The last relay protein activates a transcription factor Receptor protein 4 SIGNALING CELL Signal molecule 1 Receptor protein 2 (5) The transcription factor triggers transcription of a specific gene Plasma membrane 3 TARGET CELL Relay proteins Transcription factor (activated) (6) Translation of the mRNA produces a protein 4 NUCLEUS DNA 5 Transcription mRNA New protein 6 Translation 1. Gen merupakan materi genetik yang dapat diturunkan pada keturunannya 2. Plasmid adalah bakteri ekstrakromosomal yang dapat bereplikasi 3. Gen yang terdapat dibagian intron dapat diterjemahkan dalam proses transkripsi 4. Urutan sintesis protein dimulai dari replikasi DNA, transkripsi, dan translasi 5. Ekspresi gen diaktifkan oleh adanya perubahan lingkungan 6. Transplantasi inti sel dewasa pada sel telur, maka sel telur dapat berkembang menjadi embrio A 1. Perbanyakan sel (kloning) dapat dilakukan dengan transformasi inti sel somatik 2. Plasmid adalah protein ekstrakromosomal yang dapat bereplikasi dan dapat ditemukan pada sel hidup 3. Transkripsi untuk menghasilkan protein terjadi di sitoplasma sel 4. Gen yang terdapat dibagian ekson dapat diterjemahkan dalam proses transkripsi 5. Pengaturan gen ditentukan oleh interaksi hubungan sel ke sel 6. Stem sel dewasa dapat di kultur dan mengalami diferensiasi B KEY A 1. B 2. S (DNA) 3. S (akson) 4. B 5. B 6.B B 1. S (transplantasi) 2. S (DNA) 3. S (inti) 4. B 5. B 6.B