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Biomedical subjects

K J Beh

Publications and source records attributed to K J Beh.

At least 19 recordsLinked to original sources
PubMed1996

A tandemly repetitive DNA sequence is present at diverse locations in the genome of Ostertagia circumcincta.

A novel repetitive DNA sequence in the sheep parasitic nematode Ostertagia circumcincta was cloned and sequenced. This 1.2-kb sequence (Oc1B) was not found in the closely related cattle parasite Ostertagia ostertagi, nor in the more distantly related sheep parasites Haemonchus contortus or Trichostronylus colubriformis. Sequences similar to Oc1B were found at various genomic locations and contained a pair of 33-bp direct repeats. Oc1B also contained a single copy of a 218-bp sequence (designated OcREP) which was present in 100 to 200 copies in the O. circumcincta genome and mostly organized in distinctive tandem arrays. The dual organizational pattern of OcREP as both a satellite-like sequence as well as interspersed as single copies amongst dissimilar sequences adds to the growing evidence for the fluidity of the parasitic nematode genome, and of eukaryotic genomes in general.

Animals↗
PubMed1994

Characterization of a tandemly repetitive DNA sequence from Haemonchus contortus.

Genomic DNA from the sheep parasitic nematode Haemonchus contortus was shotgun cloned in the plasmid vector pUC18. Recombinants which gave the strongest hybridization signals to 32P-radiolabelled genomic DNA were selected as representatives of the repetitive component of the parasite DNA. One repetitive sequence which hybridized only with DNA from H. contortus and not with DNA from two other sheep nematodes (Trichostrongylus colubriformis and Ostertagia circumcincta) was further characterized by sequencing and dot blot analysis. A related repeat was found in the closely related species Haemonchus placei. Experiments to determine the genomic organization of the repeat showed that it existed in a multi-copy number tandem array. This is the first report on the characterization of repetitive DNA in sheep parasite nematodes.

Animals↗
PubMed1994

A middle-repetitive DNA sequence element in the sheep parasitic nematode, Trichostrongylus colubriformis.

A novel repetitive DNA sequence in the sheep parasitic nematode Trichostrongylus colubriformis was cloned and sequenced. A 1.1 kb repetitive sequence (Tc15) which hybridized with DNA from T. colubriformis but not with DNA from two other parasitic nematodes, Haemonchus contortus and Ostertagia circumcincta, or sheep was further characterized. Southern blot analysis showed that the repeat hybridized to a range of fragments in restriction digested T. colubriformis DNA and existed in multiple copy number tandem arrays. However, to define clearly the repetitive monomeric unit further screening of phagemid libraries containing BamH I restriction fragments using a subclone of Tc15 as a probe was carried out. Restriction map and sequence data were compiled for 3 clones containing a 145 bp highly repetitive sequence (designated TcREP) which shared homology with the original pTc15 clone. TcREP hybridized to a tandemly repeating sequence monomer of 145 bp in T. colubriformis DNA which was cloned from various genetic environments in the T. colubriformis genome. TcREP homologous sequences were also found in the genomes of two other species of the same genus (Trichostrongylus axei and Trichostrongylus vitrinus) but not in a fourth species (Trichostrongylus rugatus).

Animals↗
PubMed1994

Cellular inflammatory responses in skin of sheep selected for resistance or susceptibility to fleece rot and fly strike.

Leucocyte populations were examined in normal and inflamed skin of sheep bred for resistance (R) or susceptibility (S) to bacterial fleece rot and the common sequela, body strike caused by the dipteran parasite Lucilia cuprina. No differences between R and S lines were found in numbers of neutrophils accumulating in acute inflammatory lesions induced by activated complement, leukotriene B4, interleukin (IL)-1 beta, tumour necrosis factor-alpha, IL-8 or endotoxin from Pseudomonas aeruginosa. T19+ (alpha gamma delta T cell subset) lymphocytes and eosinophils were more prevalent in skin of sheep from the S line whereas IgE+ cells were more prevalent in skin of sheep from the R line. In an unrelated population of sheep, animals with low fleece rot scores had more intense neutrophil migration into inflammatory lesions induced by all the mediators examined than did animals with high fleece rot scores. IgE+ cells were more prevalent in animals with low fleece rot scores, although in contrast to R and S lines, T19+ cells tended to be elevated in this group of animals. The results suggest that defence mechanisms associated with IgE+ cells in skin may play an important role in resistance to fleece rot and fly strike.

Animals↗
PubMed1994

Expression of a recombinant sheep IgE gene.

An ovine cosmid library was screened with a bovine C epsilon gene probe. IgE and IgA constant region genes (C epsilon and C alpha, respectively) were isolated within a single 26 kb cosmid insert. The C epsilon gene was cloned into an expression vector which contained the rearranged VDJ genomic segment encoding an anti-(4-hydroxy-3-iodo-5-nitrophenyl) acetic acid (anti-NIP) VH chain. This chimeric construct was transfected into murine hybridoma cells producing L chain with anti-NIP antibody specificity and stable transfectomas secreting chimeric ovine-murine IgE anti-NIP antibodies were obtained. Chimeric antibody, affinity purified from culture supernatants of transfected cells on a NIP-sepharose column, was characterised by SDS-PAGE and shown to contain H and L chains of expected size. These experiments demonstrated the power of advanced molecular genetic techniques to generate immunological reagents against low abundance immunoglobulin isotypes thus facilitating monoclonal antibody production for further immunological studies.

Animals↗
PubMed1993

Organisation of the ovine immunoglobulin C epsilon gene locus: evidence for a deletion 5' of the gene.

A cosmid clone containing the ovine and C epsilon and C alpha immunoglobulin heavy chain genes was isolated and characterised. Restriction mapping and sequence analysis showed a high degree of similarity between the bovine and ovine C epsilon loci. Restriction fragment length polymorphism (RFLP) analysis of sheep genomic DNA revealed Mendelian inheritance of polymorphisms with identical variation in allele size for various restriction enzymes. This identical variation suggested that a deletion of approximately 100 bp existed at the 5' end of the smaller alleles.

Alleles↗
PubMed1993

A search for associations between major histocompatibility complex restriction fragment length polymorphism bands and resistance to Haemonchus contortus infection in sheep.

Polymorphic bands were detected within the DQB and DRB regions of the ovine major histocompatibility complex by probing TaqI digested DNA from three large sheep half-sib families derived from a highly resistant ram. All animals were phenotypically assessed for Haemonchus contortus resistance by faecal egg counts and associations with RFLP bands and haplotypes were estimated using mixed model, best linear unbiased prediction statistical methods. Although the highly resistant sire was homozygous at the MHC, no significant associations were found between any band or haplotype and faecal egg count.

Animals↗
PubMed1992

Analysis of immunoglobulin light chain loci in sheep.

A sheep kappa cDNA probe was isolated, characterized by sequence analysis and shown to have significant sequence identity to other kappa light chains. This probe and a sheep lambda light chain probe were used to estimate the extent of various sheep immunoglobulin light chain gene loci by Southern blot analysis of genomic DNA. The results showed that the sheep has a single hybridizing kappa constant gene and three to five kappa V segment bands. Segregation of three polymorphic bands at the lambda C locus indicated that they were products of separate C segments. Restriction fragment pattern variations were obtained using light chain probes on various sheep breeds, but no pattern or individual band was characteristic for a particular breed.

Amino Acid Sequence↗