► EXPERIMENTAL FIGURE 10-30 Banding on Drosophila polytene salivary gland chromosomes and in situ hybridization are used together to localize gene sequences.
(a) In this light micrograph of Drosophila melanogaster larval salivary gland chromosomes, four chromosomes can be observed (X, 2, 3, and 4), with a total of approximately 5000 distinguishable bands. The centromeres of all four chromosomes often appear fused at the chromocenter. The tips of chromosomes 2 and 3 are labeled (L = left arm; R = right arm), as is the tip of the X chromosome. (b) A particular DNA sequence can be mapped on Drosophila salivary gland chromosomes by in situ hybridization. This photomicrograph shows a portion of a chromosome that was hybridized with a cloned DNA sequence labeled with biotin-derivatized nucleotides. Hybridization is detected with the biotin-binding protein avidin that is covalently bound to the enzyme alkaline phosphatase. On addition of a soluble substrate, the enzyme catalyzes a reaction that results in formation of an insoluble colored precipitate at the site of hybridization (asterisk). Since the very reproducible banding patterns are characteristic of each Drosophila polytene chromosome, the hybridized sequence can be located on a particular chromosome. The numbers indicate major bands. Bands between those indicated are designated with numbers and letters (not shown). [Part (a) courtesy of J. Gall; part (b) courtesy of F Pignoni.]
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