Chapter3GeneticAnalysisofVirus精编.ppt

2017-4-10 1 Chapter 3 Genetic Analysis of  Virus Section 1 、 T4 phage Section 2 、λphage Section 3 、Retrovirus 2017-4-10 2 一、T4噬菌体基因组的特点 Replication cycle results in the cell lysis structurally complex (fig 9.5b) Genome of T4 ds DNA, linear, 168,903bp encodes over 250 different proteins have a unique base, 5-hydroxymethylcytosine （5-羟甲基胞嘧啶）instead of cytosine The glucosylated DNA is resistant to virtually all restriction endonucleases of the host 2017-4-10 3 一、T4噬菌体基因组的特点 T4 DNA is circularly permuted （变成环状，环化） Cut from a circular, but in each case the cut was at a different place Terminal repeats of 3000-6000bp DNA in each virion has repeated sequences at each end A headful mechanism (头部满装机制） -the packaging mechanism involves cutting DNA from a long DNA molecule containing several genome equivalents linked end-to-end (concatemer 连环体，串联体) the concatemer is cut after the head is full, cut not a specific sequence( irrespective of the sequence ) 2017-4-10 4  什么是环状排列和末端臃余？ 环状排列是指T4phage DNA分子的核苷酸排列顺序虽然是不变的,可是每一个phage从哪一个核苷酸开始则有种种可能。 末端臃余是指T4phage DNA分子的两端有相同核苷酸序列的重复。 2017-4-10 5 噬菌体T2的基因组是200kb,末端多余的碱基约1－2kb，为其基因组的0.5-3 %。 噬菌体T4的基因组是169kb,末端多余的碱基约3.38kb，为其基因组的2 %。 2017-4-10 6 末端臃余和环状排列的形成机制 1 phageDNA在寄主中合成时，形成1个包括多个单位长度的、连在一起的dsDNA分子： abcdefgabcdefgabcdefgabcdefg…... 2 Phage头部蛋白包装DNA时，被切割的DNA长度总是固定的，而不管序列如何，比完整的基因组长一些，形成了具有可变序列的T4DNA分子。对于1个phage来讲其基因组是末端臃余，对于群体phage来讲则为环状排列。 abcdefgab cdefgabcd efgabcdef g…... abcdefgab， cdefgabcd ， efgabcdef， 2017-4-10 7 2017-4-10 8 The packaging Mechanism of T4 DNA-a Headful Mechanism Figure 09-13 Generation in T4 of virus length DNA molecules with permuted sequences by a cutting enzyme, which cuts off constant lengths of DNA irrespective of the sequence. Left: nearly replicated copies of infecting T4 genome, arrow, recombination (Section 10.5); Middle: arrows, sites of enzyme attack; Right: molecules generated. 2017-4-10 9 Time course of events in T4 infection