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By Edward A. Birge

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Leading strand n' i n ft-dnaC ~ I / ,.. DNA polymerase I - --~ e-- ~ Primer 1, ... - ~)~ '\'1ti. d~aBPrimase DNA 0 polymerase III holoenzyme . Jl . l .... o~~wn . '- rNMP dNMP ,• i<'igure 1-12. DNA replication. (A) The primer RNA is laid down by a primosome complex of the dnaG and dnaB proteins. The DNA polymerase III holoenzyme complex carries out the actual synthesis of the bulk of the DNA. The individual RNA primers are removed by DNA polymerase I, and the resulting nicks are sealed by DNA ligase.

When the four reaction mixtures are subjected to electrophoresis, a ladder of individual bands is produced (Fig. 2-3). Because each lane on the gel corresponds to fragments ending in a particular base, the DNA sequence is determined by beginning with the smallest band and reading up the gel, noting which lane has the next smallest fragment. DEOXYCYTOSINE 6 NHz DIDEOXYCYTOSINE 0~ H H N~O~H2 0H H'1----1H H H + Figure 2-2. A dideoxy base compared with the normal deoxy base. The arrow indicates the position of the missing oxygen atom.

The original mRNA transcript in eukaryotic cells is longer than the final processed version and must be processed to remove intervening sequences, as in the case of the tRNA molecules. Once again the RNA splicing reaction occurs at specific sites characteristic of each type of mRNA. , phage T4, discussed in Chapter 4). Translation of the Genetic Message The lack of a physical boundary between the bacterial nucleoid and the cytoplasm means that translation can proceed at the same time as transcription.

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