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Biomedical subjects

W Bains

Publications and source records attributed to W Bains.

15 recordsLinked to original sources

Local sequence dependence of rate of base replacement in mammals.

I have analysed the local sequence context of base replacement changes in 78 processed pseudogenes. Transversions occur more often than transitions in a ratio of 3.37 to 1, and G:C is replaced 1.4 times more frequently than A:T. In addition, the bases to the 5' and 3' of the mutating base also influence the rate at which bases change, purine:pyrimidine and pyrimidine:purine pairs changing 1.2 times as fast as purine:purine and pyrimidine:pyrimidine pairs. I discuss implications of this for the mechanism of DNA polymerization in mammals.

Animals

Hybridization methods for DNA sequencing.

I have conducted a general analysis of the practicability of using oligonucleotide hybridization to sequence DNA. Any DNA sequence may be sequenced by hybridization with a complete panel of oligonucleotides. However, sequencing DNA segments over 2 kb long requires an unrealistic number of hybridization reactions. The optimal protocol is to hybridize 7-mer or 8-mer mixed oligonucleotide probes to immobilized DNA fragments 80 bp long: should this prove impractical, hybridization of labeled 270-bp fragments to immobilized mixed 10-mers is a potential alternative. Both protocols require no more experiments to sequence large regions of DNA than conventional m13-based sequencing and are much easier to automate, thus reducing the requirements for skilled personnel. In the ideal case, hybridization sequencing reduces the number of experiments required to sequence megabase DNA by 90%.

Base Sequence

Characterization of an unusual human histone H3.3 pseudogene.

The analysis of a genomic loci containing human histone H3.3 processed pseudogenes, has revealed two regions that are unusually rich in other retroposons. At one of the loci the H3.3 pseudogene is itself interrupted by 2 Alu repetitive sequences. The characterization of these two recently transposed Alus provides confirmation of the "multiple origin" hypothesis of these repetitive elements. The unusual occurrence of 3 different types of retroposons in a small region suggests that there may be particular chromosomal regions that are hot spots for retroposon insertion.

Base Sequence

Simplified format for DNA probe-based tests.

The approach I describe to using DNA probes in diagnostic tests is simpler than most existing formats. DNA in a sample is labeled by chemical reaction with bisulfite and methylamine to generate a sulfonated derivative. The DNA need not be purified to do this. The labeled sample is then incubated with an unlabeled, purified probe DNA, which is immobilized to a solid support. The amount of label remaining on the solid support after washing is detected by a monoclonal antibody that recognizes modified cytosines. The intensity of the signal depends on the amount of target DNA in the sample. Detection limits depend on the amount of immobilized DNA and on the degree of physical entrapment of the labeled DNA in sample material, but can be as low as 5 pg. This format is well suited to automation for use with existing robotic enzyme immunoassay procedures.

DNA Probes

Repetition structure of mammalian nuclear DNA.

We have used Fragmentation Sequencing logic to analyse the repetition structure of several large human genomic genes. The method, based on a proposed laboratory scheme for DNA sequencing, detects short sequences which are repeated near, but not necessarily adjacent, to each other (cryptically simple DNA). We find a low frequency of such repeats. There is a slight excess of such repeats in introns over exons, and a slight but significant excess in genomic DNA over random DNA, confirming that cryptically simple sequences are over-represented in the genome. The analysis suggests that Fragmentation Sequencing will be a suitable method for sequencing large mammalian genes.

Animals

Similarity and divergence among rodent repetitive DNA sequences.

We have analyzed the sequence of 63 B1 and 71 B2 repetitive elements from published data base sequences. The sequences conform to previously published consensus sequences, but are not identical to them. The B2 sequences show seven regions of high variability between family members, which we show points to the B2 family containing subfamilies; no similar evidence is found for subfamilies of the B1 family. The comparisons show no evidence for the emergence of species-specific variants of B1 or B2 sequences since the separation of murine and hamster lines of descent, nor of their concerted evolution within species in the last 10 million years.

Animals

MULTAN: a program to align multiple DNA sequences.

I describe a computer program which can align a large number of nucleic acid sequences with one another. The program uses an heuristic, iterative algorithm which has been tested extensively, and is found to produce useful alignments of a variety of sequence families. The algorithm is fast enough to be practical for the analysis of large number of sequences, and is implemented in a program which contains a variety of other functions to facilitate the analysis of the aligned result.

Animals