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Next we look at the colinearity of the gene and the polypeptide. Benzer s work established that the gene was a linear entity, as Watson and Crick had proposed. However, Benzer could not demonstrate the colinearity of the gene and its protein product. To do this, it is necessary to show that for every mutational change in the DNA, a corresponding change takes place in the protein product of the gene. Colinearity would be established by showing that nucleotide and amino acid changes occurred in a linear fashion and in the same order in the protein and in the cistron. Ideally, Benzer himself might have solved the colinearity issue. He was halfway there, with his 350 or so isolated mutations of phage T4. However, Benzer did not have a protein product to analyze; no mutant protein had been isolated from rII mutants. In the midst of competition to nd just the right system, Charles Yanofsky of Stanford University and his colleagues emerged in the mid-1960s with the required proof, showing that the order of a polypeptide s amino acids corresponded to the nucleotide sequence in the gene that speci ed it. Yanofsky s success rested with his choice of an amenable system, one using the enzymes from a biochemical pathway. Yanofsky did his research on the tryptophan biosynthetic pathway in E. coli. The last enzyme in the pathway, tryptophan synthetase, catalyzes the reaction of indole3-glycerol-phosphate plus serine to tryptophan and

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Selecting an Operating Point for a BJT The family of curves shown for the collector i-v characteristic in Figure 99(b) re ects the dependence of the collector current on the base current For each value of the base current, iB , there exists a corresponding iC -vCE curve Thus, by appropriately selecting the base current and collector current (or collector-emitter

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3-phosphoglyceraldehyde. The enzyme itself is made of four subunits speci ed by two separate cistrons, with each polypeptide present twice. Yanofsky and his colleagues concentrated on the A subunit. They mapped A-cistron mutations with transduction (see chapter 7) using the transducing phage P1.They rst tested each new mutant against a series of deletion mutants to establish the region where the mutation was. Then they crossed mutants for a particular region among themselves to establish relative positions and distances. The protein products of the bacterial genes were isolated using electrophoresis and chromatography to establish the ngerprint patterns of the proteins (see chapter 11). Assuming a single mutation, a comparison of the mutant and the wild-type ngerprints would show a difference of just one polypeptide spot ( g. 12.10), avoiding the need to sequence the entire protein.The mutant amino acid was

By appropriate choice of IBB , RC and VCC , the desired Q point may be selected IC C B IBB IB + VBE _ + VCE _ VCC E RC

wild-type polypeptide digests. The single spot that differs in the mutant can be isolated and sequenced, eliminating the need to sequence the whole protein.

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voltage), we can determine the operating point, or Q point, of the transistor The Q point of a device is de ned in terms of the quiescent (or idle) currents and voltages that are present at the terminals of the device when DC supplies are connected to it The circuit of Figure 912 illustrates an ideal DC bias circuit, used to set the Q point of the BJT in the approximate center of the collector characteristic The circuit shown in Figure 912 is not a practical DC bias circuit for a BJT ampli er, but it is very useful for the purpose of introducing the relevant concepts A practical bias circuit is discussed later in this section Applying KVL around the base-emitter and collector circuits, we obtain the following equations: IB = IBB and VCE = VCC IC RC which can be rewritten as VCC VCE IC = RC RC (94) (93)

(95)

The McGraw Hill Companies, 2001

Note the similarity of equation 95 to the load-line curves of s 3 and 8 Equation 95 represents a line that intersects the IC axis at IC = VCC /RC and the VCE axis at VCE = VCC The slope of the load line is 1/RC Since the base current, IB , is equal to the source current, IBB , the operating point may be determined by noting that the load line intersects the entire collector family of curves The intersection point at the curve that corresponds to the base current IB = IBB constitutes the operating, or Q, point The load line corresponding to the circuit of Figure 912 is shown in Figure 913, superimposed on the collector curves for the 2N3904 transistor (data sheets for the 2N3904 transistor are included in the CD-ROM that accompanies the book) In Figure 913, VCC = 15 V, VCC /RC = 40 mA, and IBB = 150 A; thus, the Q point is determined by the intersection of the load line with the IC -VCE curve corresponding to a base current of 150 A

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convert tiff to pdf c# itextsharp

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