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Plant Breeding New characteristics : Better qualities / Higher quantities Environmental effects : Variation Genetic effects : More stable Genetic improvement : Hybridization / Selection Genetic engineering Plant Molecular Biology Plant biology and Molecular biology Recombinant DNA techniques Plant gene analysis Isolation / Identification / Characterization Gene expression /Product analysis Transformation Chromosome Chromosome : Polynucleotide chain Nucleosome = DNA + Protein Heritable property Eukaryote: coding sequence non-coding sequence Nucleotide Basic units of nucleic acid :DNA / RNA Nucleotide: base / phosphate deoxyribose / ribose sugar Purine : Adenine / Guanine Pyrimidine : Cytosine / Thymine / Uracil Bases Nucleic acid Sugar-phosphate backbone 5’ --> 3’ phosphodiester bond Double helix / helix Complementary bases AT, AU (2 H bonds) CG (3 H bonds) DNA Unbranched polymer (nanotech: branched?) 4 subunits (dNTPs: dATP, dCTP, dGTP and dTTP) Genetic information Other function Energy carrier Signaling molecule Coenzyme Gene structure • Structural sequence : Genetic code • Regulatory sequence : Gene expression control (promoter, terminator, enhancer) Prokaryotic gene Polycistronic mRNA (found in most prokaryotes) information / code for > 1 protein uninterrupted stretch of DNA no intron copied without alteration Prokaryotic gene RNA polymerase interact loosely with chromosome slide along DNA chain bind tightly at promoter separate at terminator Prokaryotic gene Prokaryotic gene Promoter Consensus sequence centered at -35 and -10 Prokaryotic gene Terminator Specific sequence Stop signal for RNA polymerase A run of U residues downstream of a self complementary hairpin / stem-loop structure Prokaryotic gene Terminator Eukaryotic gene mostly Monocistronic mRNA : code for single protein Exon / Intron : DNA and primary transcript RNA processing for mature transcript 5’ capping RNA splicing 3’ polyadenylation Eukaryotic gene intron RNA processing stability membrane transport translation efficiency Alternative splicing : > 1 protein Genetic recombination during evolution Central Dogma Transfer of genetic information Replication Semiconservative Local separation Template reading 3’ --> 5’ direction dNTPs substrates DNA polymerase enzyme RNA primer Replication Replication accuracy Mistake less than 1/109 nct added Errors result in mutation : silent mutation loss of mutation improved / novel phenotypes Transcription At specific regions / From DNA to RNA Transcription / Transcribe / Transcript Changes in composition : properties : function : Transcription Types / Function of RNA mRNA rRNA and tRNA snRNA : splicing, telomere maintenance RNA enzyme / ribozyme : catalytic activity RNA genome : genetic material (ss / ds) RNA gene : RNA as final product Transcription DNA template (sense/antisense, +/- , template/coding) RNA polymerase (I, II, III) NTPs substrates Transcription factor Transcription activating protein Transcription Transcription Active genes: may be transcribed 1000X in each cell generation A transcript: may be translated for thousands copies Translation From polynucleotide to polypeptide Basic unit: amino acid Compositions: mRNA ribosome tRNA:amino acid other proteins mRNA as template Genetic code / Codon triplet 64 possible codons 20 amino acids 61 + 3 stop codons (UAA UAG and UGA) Degeneracy / Degenerate codons several codons for 1 amino acid except for Met and Tryp tRNA as aa carrier 70-90 nt long Amino acid attaches to the 3’ CCA Anticodon on tRNA with complementary bases to codon on mRNA Ribosome 2 subunits rRNA + 50 rProteins tRNA-mRNA lineup in correct order ss - mRNA / t RNA ls - peptide bond formation Translation Translation direction: template reading 5’ --> 3’ Tl start signal: purine-rich sequence 5’ cap AUG as tl start site Methionine as first amino acid Genetic code Template: mRNA Amino acid carrier: tRNA 31 tRNA molecules (euka) for 20 amino acids several t RNA for 1 aa Genetic code 31 tRNAs for 61 codons Some tRNA - mRNA complete matching Some tRNA - mRNA required only first 2 nt 3rd position: mismatch *Wobble* Genetic code Universal codes : conserved among organisms with minor exception: codon usage preferred codons for some amino acid due to available tRNA Translation Transcription-Translation