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C W Schmid

Publications and source records attributed to C W Schmid.

88 records · Page 5Linked to original sources

Fractionation of the sequence organization of human DNA by preparative HgCl2/Cs2SO4 density gradients.

Human DNA has been fractionated on preparative HgCl2/Cs2SO4 density gradients. Repetitious sequences are found in DNA components of all base compositions but the (G + C)-rich sequences are enriched for repetitive sequences in (A + T)-rich components are tandemly arranged; those in (G + C)-rich components tend to be interspersed with single copy sequences. It is concluded that the interspersed repetitious sequences have high (G + C) compositions.

Base Sequence↗

Intramolecular base composition heterogeneity of human DNA.

The intramolecular base composition heterogeneity of human DNA has been investigated by electron microscopic observations of partially denatured structures and by equilibrium solution thermal denaturation techniques. DNA sequences having an average length of less than 2000 base pairs are found to be heterogeneous in base composition. These heterogeneous sequences occupy a minimum of 67 to 81% of the human genome.

Base Sequence↗

The effect of silver ion binding and pH on the buoyant density of DNA and its use in fractionating heterogeneous DNA.

1. The effect of pH on the buoyant density of the complexes of Ag+ with DNA has been studied using 3H-labeled human DNA and several bacterial DNAs to determine the conditions necessary for the maximum resolution of compositional heterogeneity. In neutral CS2SO4 density gradients, Ag+ complexes with (G - C)-rich components are always denser than those with (A - T)-rich components, since (G - C)rich DNAs have a larger affinity for Ag+ than (A - T)-rich DNAs and their complexes are denser than (A - T)-rich complexes. In alkaline (pH greater than 9) CS2SO4 gradients, the buoyant density of the Ag+ - DNA complex is not a simple function of base composition. The Ag+ affinity of (A - T)-rich DNA is larger than that of (G - C)-rich DNA but the density of a (G - C)-rich complex is larger. Thus the ordering of the buoyant density changes depends on the amount of added Ag+. 2. The problem of resolving the density heterogeneity within a tracer DNA, and minor components of DNA, is explored and useful fractionation techniques are developed.

Binding Sites↗

Inverted repeat sequences in the Drosophila genome.

The properties of inverted repeat (foldback) sequences in Drosophila melanogaster DNA have been studied by HAP chromatography and electron microscope methods. Electron microscope observations show that there is a broad distribution of lengths of the duplex regions of the inverted repeats from very short to greater than 15 kb, with number and weight average values of 1.35 kb and 5.0 kb respectively. About 20% of the inverted repeats are separated by a single-strand spacer with lengths too short to observe, but the other 80% have spacers, P, with lengths ranging from 0.5 kb to greater than 30 kb. The number average and weight average spacer lengths for the total sample are 2.7 kb and 6.1 kb. With respect to the lengths of the spacers, P, between inverted repeats, the Drosophila genome differs from that of most organisms which have been studied where the spacers P are mostly too short to be measured. EM and HAP studies suggest that the average center-to-center spacing between sets of inverted repeats is 40-80 kb. The HAP studies show that there is a broad range of thermal stabilities for the duplexes formed by reassociation of inverted repeat sequences. Kinetic analysis shows that all of the frequency components of the Drosophila genome are present in the inverted repeats, the loops P, and the flanking sequences. There is a somewhat larger proportion of middle repetitive DNA in those inverted repeat duplexes which are resistant to digestion by Mung Bean Endonuclease I. These enzyme resistant duplexes comprise about 3% of the entire genome. It is estimated that there are approximately 2000-4000 inverted repeat pairs in the entire genome.

Animals↗

Interspersion of repetitive and nonrepetitive DNA sequences in the Drosophila melanogaster genome.

Cot analysis shows that the haploid Drosophila genome contains 12 percent rapidly reassociating, highly reiterated DNA, 12 percent middle repetitive DNA with an average reiteration frequency of 70, and 70 percent single-copy DNA. The distribution of the middle repetitive sequences in the genome has been studied by an examination in the electron microscope of the structures obtained when middle repetitive sequences present on large DNA strands reassociate and by the hydroxyapatite binding methods developed by Davidson et al. (1973). At least one third by weight of the middle repetitive sequences are interspersed in single-copy sequences. These interspersed middle repetitive sequences have a fairly uniform distribution of lengths from less than 0.5 to 13 kb, with a number average value of 5.6 kb. The average distance between middle repetitive sequences is greater than 13 kb. The data do not exclude the possibility that essentially all of the middle repetitive sequences have the interspersion pattern described above; however, it is possible that some of the middle repetitive sequences of Drosophila are clustered in stretches of length much greater than 13 kb. The interspersion pattern of the middle repetitive sequences in Drosophila is quite different from that which occurs in the sea urchin, in Xenopus, in rat, and probably many other higher eucaryotes.

Animals↗

Sequence organization of the human genome.

The organization of three sequence classes--single copy, repetitive, and inverted repeated sequences--within the human genome has been studied by renaturation techniques, hydroxylapatite binding methods, and DNA hyperchromism. Repetitive sequence classes are distributed throughout 80% or more of the genome. Slightly more than half of the genome consists of short single copy sequences, with a length of about 2 kb interpersed with repetitive sequences. The average length of the repetitive sequences is also small and approximates the length of these sequences found in other organisms. The sequence organization of the human genome therefore resembles the sequence organization found in Xenopus and sea urchin. The inverted repeats are essentially randomly positioned with respect to both sequence class and sequence arrangement, so that all three sequence classes are found to be mutally interspersed in a portion of the genome.

Base Sequence↗

Isolation and chromosomal localization of highly repeated DNA sequences in Drosophila melanogaster.

The nuclear DNA of D. melanogaster contains DNA sequences that are repeated between ten and a hundred times more often than the next class of redundant DNA. This DNA, as a renatured duplex isolated on the basis of its renaturation kinetics, has a buoyant density of 1.691 (g/ml). In its native state it bands within the unique nuclear DNA peak (rho = 1.701). These sequences have been localized by "in situ" hybridization in the chromocenter of the chromosomes of the salivary gland. The properties of centromeres are discussed in terms of the occurrence of repeated sequences at this locus.

Animals↗

A transposon-like element in human DNA.

Mobile genetic elements have been reported in prokaryotes, plants, yeast and Drosophila. The only transposon-like sequences reported for mammalian organisms are closely related to retroviruses, although undoubtedly other transposon families exist within the mammalian genome. Although mobile genetic elements can only be identified as such if their mobility can be demonstrated in existing populations, transposon and transposon-like elements share several common biochemical and structural features. Here we demonstrate that a repetitive human sequence has many of the diagnostic features of transposable elements. This 2.3-kilobase (kb) transposon-like element contains two flanking long terminal repeat (LTR)-like 350-base pair (bp) repetitive sequences, each of which begins with the sequence 5' TG... and ends with ...CA 3'. The transposon-like element is bounded by 5-bp direct repeats. Discrete-length polyadenylated transcripts from HeLa cells are homologous to the transposon-like element. Members of this transposon-like family are found in extrachromosomal circular DNA molecules.

Animals↗