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Biomedical subjects

A G Mackinlay

Publications and source records attributed to A G Mackinlay.

At least 19 recordsLinked to original sources
PubMed1996

Detection and characterization of two novel hypervariable microsatellite repeat regions within intron 2 of the alpha-globin gene of the bivalve mollusc Anadara trapezia.

Synthetic oligonucleotide primers based on cDNA sequence were used to amplify the region spanning intron 2 of the alpha-globin gene of the bivalve mollusc Anadara trapezia. Amplification of this region from individual clams showed highly polymorphic patterns. The sequence of this intron was found to include a number of mono- [d(T)n and d(C)n], di- [d(CA)n and d(CT)n] and tetranucleotide d(CTGT)n repeats which were found to be polymorphic with respect to the types and numbers of repeats present. Two separate repeat-containing polymorphic regions were located near each end of this intron. The repeat at the 3' end consisted of an unusual example of a d(T)n polymorphism at the position of the polypyrimidine tract usually involved in intron splicing. Thirteen individual cloned intron 2 sequences, derived by PCR amplification from pooled genomic DNA, were sequenced without finding two identical sequences. All of the sequenced clones contained microsatellite sequences.

Animals↗
PubMed1995

Characterisation of a second, apparently inactive, copy of the bovine beta-lactoglobulin gene.

A bovine cosmid clone was isolated which contains the previously characterised beta-lactoglobulin gene and, in addition, a related sequence which appears to be a beta-lactoglobulin pseudogene. The total length of the pseudogene, as determined by DNA sequencing, is 4.8 kb, similar to that of beta-lactoglobulin. Both genes are in the same orientation and are separated by approximately 14 kb intergenic sequence. Although most of introns I-V are extremely divergent, the exon sequences are clearly related, exons I-V exhibiting nucleotide sequence similarities in the range 60-87%. Exons VI and VII, together with the final intron, comprise a region of sequence extending over 730 bp, which displays 92.5% identity to the corresponding beta-lactoglobulin sequence. It is suggested that this is the result of a recent gene conversion event involving conversion of the pseudogene by the authentic beta-lactoglobulin gene. Identification of the new lactoglobulin sequence as a pseudogene is based on the occurrence of a stop codon in exon V. Comparison of the inferred translation product encoded by the pseudogene before its mutational inactivation, with the sequences of equine and feline beta-lactoglobulins I and II, indicates that the bovine pseudogene is more closely related to these type-II lactoglobulin sequences than to the type-I sequences.

Amino Acid Sequence↗
PubMed1994

The organization of the beta-globin gene of the bivalve mollusc Anadara trapezia and its evolutionary relationship to other invertebrate and vertebrate globin genes.

Three pairs of oligonucleotide primers based on partial DNA and amino acid sequences were used in a combination of PCR experiments to amplify the beta-globin gene of the bivalve mollusc Anadara trapezia. The sequence of 2,139 bp presented contains the whole of the beta-globin gene with the exception of the 5' flanking sequence. This gene possesses the three-exon-and-two-intron gene structure typical of vertebrate globin genes but the lengths of the introns (762 bp and 690 bp, respectively) are only approximately half the size of those present in a beta-variant gene previously characterized from this organism. The encoded amino acid sequence shows two changes when compared to the previously published amino acid sequence.

Amino Acid Sequence↗
PubMed1993

PCR amplification of partial mRNA sequences encoding the alpha- and beta-globin chains of the bivalve mollusc Anadara trapezia: correction of the C-terminal amino acid sequence of the alpha-chain.

1. PCR was carried out on cDNA which had been synthesized from poly(A)+ RNA extracted from A. trapezia red blood cells. 2. A 348 bp product encoding the C-terminal region of the alpha-globin chain and another product encoding a portion of the beta-globin chain were isolated. 3. The C-terminal alpha-chain sequence, encoded by the cDNA, differs from the previously published sequence and these changes improve the alignment of this chain with the other globin chains of this organism.

Amino Acid Sequence↗
PubMed1989

The CO-I and CO-II region of honeybee mitochondrial DNA: evidence for variation in insect mitochondrial evolutionary rates.

The sequence of a region of honeybee (Apis mellifera ligustica) mitochondrial DNA, which contains the genes for cytochrome c oxidase subunits I and II (CO-I and CO-II) and inferred genes for tRNA(Asp), tRNA(Leu)UUR, tRNA(Lys), and tRNA(Trp), is presented. The region includes the segment previously identified as incurring a length increase in some other bee strains, including Africanized bees. The sequence information of this study and of that by Vlasak et al. shows that several shifts of tRNA genes have occurred between Apis and Drosophila, but shifts of other kinds of genes have yet to be demonstrated. The CO-I and CO-II gene sequences are both more A+T rich than are the corresponding Drosophila genes. Parsimony analyses using the mouse and Xenopus sequences as outgroups show significantly more amino acid substitutions on the branch to Apis (120) than on that to Drosophila (44), indicating a difference in the long-term evolutionary rates of hymenopteran and dipteran mtDNA.

Amino Acid Sequence↗
PubMed1988

Isolation and characterization of the bovine kappa-casein gene.

1. The bovine kappa-casein gene has been isolated as a series of overlapping lambda clones and shown to consist of five exons distributed over a total length of approximately 13 kb. Most of the mature protein-coding sequence is contained in a single large exon. 2. Approximately 65% of the gene has been sequenced together with portions of the 5'- and 3'-flanking sequences. The immediate 5'-flanking sequence contains several motifs which are characteristic of upstream regions including a TATA box, a CAAT box, a sequence similar to that recognized by transcription factor AP-1 and a purine-rich sequence resembling that found upstream in all other lactoprotein genes. Other possible regulatory sequences are found upstream of exon 4. 3. The organization of the kappa-casein gene, together with its upstream sequence, confirms previous conclusions that it is unrelated to the calcium-sensitive-casein gene family to which it is linked. Evidence is presented which supports a previous suggestion that kappa-casein and the fibrinogens are evolutionarily related. 4. Intron sequences contain several examples of the A family of the artiodactyl Alu-like repeated sequences, together with a single example of a C-family sequence. The remainders of the introns of the kappa-casein gene, compared with the repeat elements and exons, are A + T-rich. 5. Among the lambda clones isolated, representatives were found of the A and B genetic variants which can be distinguished by restriction-enzyme analysis. Several other examples of polymorphisms in the non-coding region were found.

Alleles↗
PubMed1988

Complete nucleotide sequence of the bovine beta-casein gene.

The beta-casein gene is a member of a small gene family encoding the calcium-sensitive caseins, which are specifically synthesized and secreted by the mammary gland during lactation in response to both peptide and steroid hormones. The caseins are involved in the transport of calcium phosphate in milk, which is important for bone development in the infant mammal. We report here the organization and complete DNA sequence of the 8.5 kb long bovine beta-casein gene. Comparison with the rat beta-casein gene reveals that the exons of both genes correspond exactly. The 5' flanking sequences of all Ca-sensitive casein genes are conserved within the proximal 200 bp and contain several elements that probably function as cis-acting regulatory elements, including an octamer-like motif, and SV40-type core enhancer and a sequence that appears to be common to all lactoprotein genes. The latter sequence is flanked on either side by 12 bp direct repeats. These direct repeats are themselves each part of sequences that display two-fold symmetry. The first 30 nucleotides of the 3' flanking regions in the bovine and rat beta-caseins are well conserved, indicating that they are likely to be involved in the mechanism of 3' end processing of the primary transcript.

Amino Acid Sequence↗
PubMed1988

Family studies in ornithine transcarbamylase deficiency.

Six families with at least one infant each with confirmed ornithine transcarbamylase deficiency were investigated by DNA analysis. All the affected sons had died, and no DNA had been stored. Using the restriction endonucleases MspI and Bam HI three restriction fragment length polymorphisms were detected which led to eight distinct haplotypes. Using these results and those of protein loading tests that diagnosed heterozygote (carrier) status in some family members, some carriers were detected, and prenatal diagnosis was offered to two families. In two further families no polymorphisms were found and no prenatal diagnosis was possible. In the remaining two families prenatal diagnosis was impossible because of the lack of DNA from an affected or unaffected son, or in one case from the father, of an obligate carrier. These studies emphasise the importance of preserving tissue for DNA extraction from infants dying of inborn errors of metabolism, and also show the way in which information from conventional biochemical studies can complement diagnostic tests using DNA.

Amino Acid Metabolism, Inborn Errors↗
PubMed1988

Casein gene expression in bovine mammary gland.

The objectives were to examine the rate of synthesis of casein mRNA transcripts in bovine mammary tissue at different hormonal states and to study the effects of hormonal stimuli (insulin, hydrocortisone, and prolactin) on the accumulation of casein mRNA and on the rate of protein secretion by epithelial cells from bovine mammary tissues. Total cytoplasmic RNA was extracted from mammary tissues of cows obtained by biopsy (8 mo pregnant) and upon slaughter (lactating). The relative specific activities of cytoplasmic mRNA for alpha s1-, alpha s2-, beta-, and kappa-casein were about 3.2, 4.6, 3.3, and 4.5-fold higher in tissues of lactating cows than in those of 8 mo pregnant cows. Mammary alveolar epithelial cells retained hormone-inducible milk protein gene expression for total milk protein gene expression for total milk protein secretion and for alpha s1- and beta-casein messages. Prolactin, even in the absence of insulin and hydrocortisone, induced significant amounts of milk protein mRNA. Hydrocortisone in the presence of prolactin amplified the lactogenic effects on mammary epithelium. Maximal induction of beta-casein mRNA and protein secretion occurred when all three hormones were present.

Animals↗
PubMed1988

The use of field-inversion gel electrophoresis for deletion detection in Duchenne muscular dystrophy.

Deletion is a common cause of Duchenne muscular dystrophy (DMD). Field-inversion gel electrophoresis, in conjunction with Southern blot hybridization, was used to detect large SfiI DNA fragments in the DMD locus. Two unrelated boys with DMD were found to have abnormal sized DNA fragments resulting from deletions. Some of the female relatives of these patients were also shown by this method to have deletions in the DMD locus.

Chromosome Deletion↗
PubMed1987

Genetic variation in mitochondrial DNA from some aboriginal Australians.

Mitochondrial (mt) DNAs from 17 aboriginal Australians, predominantly from the coastal region of the Northern Territory were isolated and digested with four four-base restriction endonucleases, two of which revealed variation between samples. The observed fragment patterns were used directly in parsimony analyses of phylogenetic relationships between the samples, and were also converted to estimates of the number of substitutions per nucleotide position between samples (delta), which estimates were then used in distance analyses of phylogeny. The inferred fragment patterns of the completely sequenced 'Cambridge' human mtDNA were also included in these analyses. No strong evidence of geographic variation was found, consistent with previous findings of Australian aborigines and other humans generally, although the most divergent sample was one of two from Sydney, indicating that further work is desirable. The estimate of mean difference between samples (diversity), 0.0017 +/- 0.0003 (mean +/- 95% confidence interval), is significantly lower than that reported previously for humans generally.

Australia↗
PubMed1987

Complete nucleotide sequences of bovine alpha S2- and beta-casein cDNAs: comparisons with related sequences in other species.

The nucleotide sequences corresponding to bovine alpha S2- and beta-casein mRNAs have been determined by cDNA analysis. Both sequences appear to be complete at their 5' ends. The nucleotide sequence of alpha S2-casein, when compared with the corresponding cavine A sequence, helps to define the boundaries of a large amino acid repeat (approximately 80 residues) whereas comparisons with the nucleotide sequences of rat gamma- and mouse epsilon-casein mRNAs also reveal extensive sequence similarities. An alignment of these four sequences shows that the divergence of their translated regions has been characterized by the duplication and deletion of discrete segments of sequence that probably correspond to exons. A high degree of nucleotide substitution is also found when the four sequences are compared, except for well-conserved leader-peptide and phosphorylation-site sequences and, to a lesser extent, the 5'-untranslated regions. Similar comparison of the bovine and rat beta-caseins shows that their divergence has involved a high rate of nucleotide substitution but that no major insertions or deletions of sequence have occurred. The several splice sites that have veen defined in the rat beta-casein gene are likely to have been conserved in the bovine. The contrasting evolutionary histories of the alpha- and beta-casein coding sequences correlate with the distinctive functions of these proteins in the casein micelle system in milk.

Animals↗