barcode scanning in c#.net Adenine E. coli Euglena Poliovirus 24 26 30 Uracil 22 19 25 Guanine 32 31 25 Cytosine 22 24 20 in Software

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Adenine E. coli Euglena Poliovirus 24 26 30 Uracil 22 19 25 Guanine 32 31 25 Cytosine 22 24 20
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Tamarin: Principles of Genetics, Seventh Edition
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III. Molecular Genetics
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10. Gene Expression: Transcription
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Gene Expression: Transcription
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Prokaryotic Initiation and Termination Signals for Transcription
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The DNA region that RNA polymerase associates with immediately before beginning transcription is known as the promoter. The promoter is an important part of gene expression in both prokaryotes and eukaryotes. Promoters contain the information for transcription initiation and are the major sites in which gene expression is controlled. Without the sigma factor, the core enzyme of RNA polymerase binds randomly along the DNA. Formation of the holoenzyme brings about high af nity of RNA polymerase for DNA sequences in the promoter region. Termination of transcription comes about when the polymerase enzyme recognizes a DNA region known as a terminator sequence. Let us elaborate on the various stages of transcription (in this section and in boxes 10.1 and 10.2).
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Protein DNA
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Nucleases
Promoters
The RNA polymerase molecule covers a region of about sixty base pairs of DNA. This was determined by causing the polymerase to bind to DNA and then digesting the mixture with nucleases, in a technique known as footprinting ( g. 10.5). The polymerase protects or prevents degradation of the region it covers. The undigested DNA is then isolated and its size determined. Geneticists have gained much new information about the nature of recognition regions within promoters through recombinant DNA technology and nucleotide sequencing techniques (see chapter 13). Sequencing of numerous promoters has shown that they contain common sequences. If the promoter nucleotide sequences align with each other, and each has exactly the same series of nucleotides in a given segment, we say that the sequence of that segment comprises an invariant or conserved sequence. If, however, there is some variation in the sequence, but certain nucleotides occur at a high frequency (signi cantly greater than by chance), we refer to those nucleotides as making up a consensus sequence. Surrounding a point in prokaryotic promoters about ten nucleotides before the rst transcribed base is just such a consensus sequence TATAAT. This sequence is known as a Pribnow box after one of its discoverers ( g. 10.6). The nucleotides in the Pribnow box are mostly adenines and thymines, so the region is primarily held together by only two hydrogen bonds per base pair. Since local DNA denaturation occurs during transcription by RNA polymerase (the DNA is opened to allow transcription), fewer hydrogen bonds make this process easier energetically. When the polymerase is bound at the promoter region ( g. 10.6), it is in position to begin polymerization six to eight nucleotides down from the Pribnow box.
Isolate and characterize remaining DNA
Footprinting technique. DNA in contact with a protein (e.g., RNA polymerase) is protected from nuclease degradation. The protected DNA is then isolated and characterized.
Promoter region
First base transcribed
PR T7A1 T7A2 XI74A
5 T G G C G G T G A T A A T G G T T G C A T G T 3 C C T A T A GG A T A C T T A C A G G C A T C A T G C A G T A A G A T A C A A A T CGC T A T G T A T G T T T T C A T G CC T C C A A A T CG G C T CGT A T G T T G T G T G G A A T T G C A G C TT A T A A T GG T T A C A A A T A Pribnow box
SV40
Nucleotide sequences of the promoter region and the rst base transcribed from several different genes. Lambda ( ), T7, and 174 are bacteriophages. Lac is an E. coli gene, and SV40 is an animal virus. Only the SV40 promoter has the actual consensus sequence of TATAAT. Even when other sequenced promoters not shown here are Included, no base is found 100% of the time (conserved).
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