By Gary D. Burkholder (auth.), J. Perry Gustafson, R. Appels (eds.)

A historic point of view at the examine of Chromosome constitution and serve as R. Appels department of Plant CSIRO P.O. field 1600 A.C.T. AUSTRALIA "Modern actual technological know-how offers us no version to give an explanation for the re­ duplication of the gene-string in every one telephone iteration, or to ex­ undeniable the construction of powerful amounts of particular enzymes or different brokers by way of particular genes. the suitable pairing and inter­ swap of segments by means of homologous gene-strings at meiosis additionally recommend novel actual homes of this type of matter". Stadler (1954) The very powerful impression of reductionism within the heritage of realizing chromosome constitution and serve as is obvious within the above citation from Stadler's 1954 paper, "The gene". Earlyob­ servations at the fidelity of the cytological visual appeal of chromo­ somes and their standard behaviour in mobilephone department resulted in specula­ tion on their organic significance. As genetics grew to become extra subtle within the early many years of the twentieth century the genes-on-a­ string version of chromosomes built and larger emphasis was once put on the extra dissection of those constructions. accordingly, within the 1980's the reductionist method is attaining a crest as vast areas of the genetic fabric are being sequenced.

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BURKHOLDER rearrangements or alterations ,occur in the chromosomes during the banding pretreatments? 2. What specific effects do the pretreatments have on the chromosomal proteins? 3. Which proteins are involved? 4. How does the Giemsa dye stain chromosomes? 5. How do the proteins in different regions of the chromosome modulate Giemsa staining to produce banding? Although many aspects of the mechanism of G-banding are still in doubt, partial answers to some of these questions are now known. As a result of the normal condensation of chromatin to form the compact metaphase chromosome and the additional contraction induced by the standard use of colcemid as a mitotic blocking agent, acid-fixed chromosomes are almost always uniformly-stained by Giemsa.

G- and C-banding have recently been obtained by digesting unfixed, isolated chromosomes with certain restriction endonucleases (Burkholder and Schmidt, 1986; Burkholder, 1987). Interestingly, C-bands can be directly visualized after the enzyme digestion by phase-contrast microscopy, whereas G-bands are observed only after the post-digested chromosomes are fixed with acetic-methanol. This suggests that a fixation step is required to visualize G-bands. , 1970). It is technically the simplest of all the banding methods.

This is further emphasized by studies involving BrdU substitution for thymine during DNA replication followed by staining with 33258 Hoechst (Latt, 1973). The fluorescence of Hoechst is quenched by BrdU-substituted DNA, so if DNA is substituted with BrdU during only part of the S phase, chromosome regions containing substituted and unsubstituted DNA can be identified at the subsequent mitosis. These studies have directly correlated late-replicating regions with G-bands. , 1970; Bostock and Prescott, 1971).

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