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Regulation of gene expression III. tRNA synthesis, aminoacyl activation, translation, translation inhibitors, molecular genetics of deseases and diagnostic methods Downloaded from www.pharmacy123.blogfa.com
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The genetic code Downloaded from www.pharmacy123.blogfa.com
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The structure of transfer RNA Ψ: pseudouridineD: dihydrouridine Downloaded from www.pharmacy123.blogfa.com
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Activation of aminoacids Downloaded from www.pharmacy123.blogfa.com
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Comparison of ribosomes Downloaded from www.pharmacy123.blogfa.com
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Initiation of translation 1. Peptidyl- transferase 2. EF-G (translocase) Downloaded from www.pharmacy123.blogfa.com
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Elongation 1. EF-Tu + GTP= EF-Tu-GDP + Pi EF-Ts EF-Tu + GDP Important time delay for accuracy Downloaded from www.pharmacy123.blogfa.com
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Termination Binding of releasing factor to STOP Termination Downloaded from www.pharmacy123.blogfa.com
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Translation on the surface and processing in the lumen of ER Downloaded from www.pharmacy123.blogfa.com
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The recognition of signal peptide Downloaded from www.pharmacy123.blogfa.com
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Double transmembrane domain Stop-transfer sequence Start-transfer sequence Downloaded from www.pharmacy123.blogfa.com
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Multiple transmembrane domain Downloaded from www.pharmacy123.blogfa.com
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Differences in the pro-and eucaryotic mRNA Downloaded from www.pharmacy123.blogfa.com
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The procaryotic translation Downloaded from www.pharmacy123.blogfa.com
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Regulation of translation initiation dsRNA dependent protein kinase phosphorylates eIF, this inhibits initiation Viral interferon stimulated dsRNA dependent protein kinase (initial stage of infection:-RNA, +RNA or RNA:RNA hybrid) Reticulocyte protein kinase: inhibited by hem – no hem, no protein synthesis (90% globin) Downloaded from www.pharmacy123.blogfa.com
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The effect of antibiotics on procaryotic translation Steptomycin – inhibition of initiation, misreading of the code (resistance: mutation in the S12 ribosomal protein) Neomycin, gentamycin similarly Tetracycline – inhibits a. ac. tRNA binding to ribosome Chloramphenicol – inhibits pepidyl transferase Downloaded from www.pharmacy123.blogfa.com
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Eucaryotic translation specific inhibitors Puromycin – prevents tRNA binding to ribosome and incorporates into peptide chain, cause premature termination Cycloheximide – specific for eucaryotes, chloramphenicol-like effect Diptheria (corynebacterium exotoxin) the elF2 is inactivated by ADP-ribosilation made by the toxin Downloaded from www.pharmacy123.blogfa.com
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DNA chip Green: less than average exp. red: more than average exp. black: close to average exp. Differential analysis of mRNA levels of 1800 genes on cDNA array Downloaded from www.pharmacy123.blogfa.com
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Differential analysis of protein expression pH2pH12 Downloaded from www.pharmacy123.blogfa.com
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Fluorescence-activated cell sorter Downloaded from www.pharmacy123.blogfa.com
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Sequencing I Downloaded from www.pharmacy123.blogfa.com
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Sequen cing II. Downloaded from www.pharmacy123.blogfa.com
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Sequencing III. Downloaded from www.pharmacy123.blogfa.com
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RNA interference Downloaded from www.pharmacy123.blogfa.com
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Genetic diseases 4000 diseases have genetic base 10% of hospitalised children Many chronic diseases have genetic component (diabetes melitus, athelosclerosis) Chromosomal - trisomy of 21 – Down’s syndrome (karyotype – chromosomal translocations – positional effect) Monogenic disorders (autosomal dominant – recessive – X-linked) Multifactorial dirorders (ishemic heart disease, hypertension) Downloaded from www.pharmacy123.blogfa.com
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Pedigree analysis T to A change in the -globin gene – sickle cell anemia SS homozygous AS heterozygous (prenatal) AA homozygous Downloaded from www.pharmacy123.blogfa.com
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