基因工程打印稿.pptxVIP

第五章? 噬菌体的遗传分析 Bacteriophage Genetics第一节 噬菌体的繁殖和突变型 Life cycle and mutants of phagesGenetic material in a phage一、烈性噬菌体（ virulent phage）Phage T4: Structure and life cycle ◇ one of T-even phages ◇ intricate structure ◇ genetic material is DNA, linear, about 165 kb, encode for more than 150 genes Life cycleLytic development is controlled by a cascade ( 裂解周期中基因产物的多级调节 )◆早期：由宿主RNA多聚酶转录噬菌体基因，产物主要为RNA多聚酶，δ因子或反终止因子◆中期：早期基因产物转录中期基因，产物为δ因子或反终止因子、复制酶等◆晚期：中期基因产物转录晚期基因，产物为新的噬菌体组装所需的各个组分。 二、温和噬菌体（temperate phage)Phage λ: Structure and life cycle ◇ The genome is 48514 bp ◇ linear double-stranded DNA, form a ring after the phage injected into the host cell through the sticky ends ◇ clusters of genes with related functions Life cycle: ◇ lytic cycle (裂解周期） ◇ lysogenic cycle（溶源周期）Key concepts: ◇ lysogeny（溶源性） ◇ prophage（原噬菌体）　◇ lysogenic bacteria or lysogen （溶源菌）Integration of λchromosome initiate the lysogenic cycleLysogenic bacterium are immune to further injection by the same phageThe λ repressor protein is responsible for maintaining lysogenic state and for immunity to superinfection Events that damage DNAmay induce prophages toenter the lytic cycleZygotic induction (合子诱导) Hfr（λ）×F－ Hfr（λ）×F－（λ） The λ lytic cycle is interlocked with the lysogenic cycle 三、噬菌体突变型（phage mutants）Phages are so small that they are visible only under the electron microscopeWe cannot produce a visible colony by platingThe formation of a plaque ( a clear area) phage phenotype Plaque morphology（噬菌斑形态突变型 ）Altered host range（宿主范围突变型 ）Conditional lethal （条件致死突变型 ） ◇ temperature sensitive mutations （ts） （温度敏感突变） ◇ suppressor-sensitive mutations（sus） （抑制因子敏感突变 ） Suppressor-sensitive mutationsNonsense mutation（无义突变）: 琥珀型（amber）：UAG 赭石型（ocher）：UAA 乳白型（opal）：UGA Nonsense mutation is conditional lethal mutation, because the host has nonsense suppressor Nonsense suppressor The nonsense suppressors encoded by altered tRNA genes can insert a specific amino acid at the position of stop codon. Nonsense mutati