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Biomedical subjects

G C Overton

Publications and source records attributed to G C Overton.

7 recordsLinked to original sources

Fast Fourier transform-based correlation of DNA sequences using complex plane encoding.

The detection of similarities between DNA sequences can be accomplished using the signal-processing technique of cross-correlation. An early method used the fast Fourier transform (FFT) to perform correlations on DNA sequences in O(n log n) time for any length sequence. However, this method requires many FFTs (nine), runs no faster if one sequence is much shorter than the other, and measures only global similarity, so that significant short local matches may be missed. We report that, through the use of alternative encodings of the DNA sequence in the complex plane, the number of FFTs performed can be traded off against (i) signal-to-noise ratio, and (ii) a certain degree of filtering for local similarity via k-tuple correlation. Also, when comparing probe sequences against much longer targets, the algorithm can be sped up by decomposing the target and performing multiple small FFTs in an overlap-save arrangement. Finally, by decomposing the probe sequence as well, the detection of local similarities can be further enhanced. With current advances in extremely fast hardware implementations of signal-processing operations, this approach may prove more practical than heretofore.

Algorithms

Expression of SPARC/osteonectin transcript in murine embryos and gonads.

A cDNA clone, p2-4, was isolated from mouse teratocarcinoma-derived parietal endoderm-like cells and used to analyze expression of the corresponding transcript during mouse embryogenesis. Nucleotide-sequence analysis revealed extensive homology between this clone and SPARC/osteonectin cDNA cloned from mouse parietal endoderm and bovine bone cells. The SPARC/osteonectin transcript became more abundant when embryonal carcinoma (EC) cells differentiated into parietal endoderm-like cells. In embryos, the transcript began to appear in the embryo proper on day 11 and continued to be expressed throughout the gestation period. The transcript was also present in extraembryonic membranes and placenta from days 9 and 11 onward, respectively. Thus, expression of the transcript was regulated during differentiation of EC cells and during embryogenesis. In adult mice, several non-bone tissues, including testis, also expressed the transcript. Analysis of germ-cell-deficient mice indicated that non-germ-cell components of the testis expressed the transcript. Analysis of mouse testicular cell lines further suggested that the transcript was abundant in Sertoli cells and Leydig cells. Cumulus oophorus cells that envelope the ovulated egg also expressed high levels of the transcript.

Animals

Expression of the intracisternal A-particle is elevated during differentiation of embryonal carcinoma cells.

Three cDNA clones coding for the 3' region of the intracisternal A-particle (IAP), a mouse endogenous retrovirus, were isolated during screening of a library for genes whose expression was modulated during the retinoic acid-induced differentiation of the embryonal carcinoma cell line F9 into parietal endoderm-like (PE-like) cells. In contrast to previously reported results, no IAP transcripts were detected in either F9 cells or two pluripotent cell lines tested. Instead, IAP transcripts as well as IAPs were abundant in the PE-like cells PYS-2 and F9AcCl 9 and in retinoic acid-induced F9 cells but not in the other differentiated cell types of teratocarcinoma origin which were examined. A comparison of the nucleotide sequences of the three IAP cDNA clones with a genomically integrated proviral sequence (MIA14) demonstrated heterogeneity in both length and sequence among the clones. The position of the poly(A) addition site was determined to be 15 nucleotides from the proposed poly(A) addition signal and to occur after the sequence CAGA, not CA, as previously proposed. Length heterogeneity was greatest in a region of TC repeats 80 base pairs 5' to the poly(A) addition site. Additionally, the putative TATAA box found in MIA14 was deleted in the cDNA clones and in the long terminal repeat regions from two other genomic clones examined. The heterogeneity evident among the cDNA clones further demonstrated that at least two distinct IAP genes are activated during differentiation. An analysis of the rate of transcription in isolated nuclei indicated that the activation of expression of IAP genes in PE-like cells is the result of transcriptional regulation. Together, these observations suggest that the modulation of IAP transcription is regulated autonomously rather than by the fortuitous integration of an IAP sequence adjacent to a developmentally regulated cellular gene.

Animals

Length and sequence heterogeneity of the histone gene repeat unit of the sea urchin, S. purpuratus.

Histone gene repeats in S. purpuratus are shown to be of variable length and sequence. Two recombinant plasmids containing the full-length 6.3 kb histone repeat unit are found to differer in length at two sites in the repeating structure and in the occurrence of two restriction enzyme recognition sites. Variation in repeat length is also demonstrated in the unfractionated DNA of five sea urchins and in a sample of DNA enriched for histone gene sequences by density gradient methods. The repeats in each individual are of a very limited number of major classes, which may differ from one another in overall length or in distribution and presence of particular restriction enzyme sites. Variations are found to occur at many regions of the repeat; some have been mapped specifically to spacer regions. Repeats may differ dramatically from individual to individual since there is no one type of repeat class common to all, although the absolute length differences of the repeats that are found are small.

Base Sequence

Histone gene arrangement in the sea urchin, Strongylocentrotus purpuratus.

The DNA coding for histones from Strongylocentrotus purpuratus, purified up to 100-fold with the use of Hg+2-CS2-SO4 and actinomycin-CsC1 equilibrium density gradients, has been used to study the clustering of genes coding for different histones and the size of the repeating multigene cluster. When digested with EcoRI restriction endonuclease, the histone DNA is identified in two classes of fragments with molecular weights of 1.15 X 106 and 2.8 X 106, whereas after treatment of the DNA with HindIII restriction endonuclease, histone gene sequences can be identified only in a fragment of 3.95 X 106. Treatment of the DNA with both enzymes simultaneously shows that there is a HindIII site within the smaller EcoRI fragment. Partial digests with HindIII give fragment sizes that appear to be simple multiples of a 3.95 X 106 repeat. Individual histone mRNAs all hybridize to the 3.95 X 106 fragment but only to one or the other EcoRI fragments. The evidence strongly suggests a repeating unit of 3.95 X 106 containing the genes for most, if not all, the histonrs.

Animals