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Topic:Molecular biology

Molecular biology in horses involves the study of molecular processes and genetic mechanisms that underpin equine physiology and health. This field encompasses the analysis of DNA, RNA, proteins, and other biomolecules to understand gene expression, genetic variation, and cellular functions in horses. Techniques such as genomic sequencing, gene expression profiling, and molecular diagnostics are employed to explore topics like hereditary diseases, performance traits, and immune responses in equines. This page assembles peer-reviewed research studies and scholarly articles that investigate the molecular biology of horses, focusing on genetic research, molecular techniques, and their applications in equine science.
Monoclonal antibodies to the equine CD2 T lymphocyte marker, to a pan-granulocyte/monocyte marker and to a unique pan-B lymphocyte marker.
Immunobiology    December 1, 1994   Volume 192, Issue 1-2 48-64 doi: 10.1016/S0171-2985(11)80407-9
Tumas DB, Brassfield AL, Travenor AS, Hines MT, Davis WC, McGuire TC.Murine monoclonal antibodies, HB88A, B29A and DH59B separately identify the CD2 T lymphocyte molecule, a unique pan-B lymphocyte surface marker and a pan-granulocyte/monocyte surface molecule, respectively, in the horse. Specificity was shown by two-color immunofluorescent flow cytometry and immunofluorescent microscopy. MAb HB88A reacted with a 52 kDa pan-T lymphocyte molecule present on 75% +/- 7 of peripheral blood lymphocytes (PBL) (n = 15 horses). It also reacted with lymphocytes restricted to T lymphocyte dependent areas of lymph node and spleen. Specificity of mAb HB88A to CD2 was demon...
Sequence analyses of the p24 gene of Borna disease virus in naturally infected horse, donkey and sheep.
Virus research    December 1, 1994   Volume 34, Issue 3 281-289 doi: 10.1016/0168-1702(94)90128-7
Binz T, Lebelt J, Niemann H, Hagenau K.By reverse transcriptase/PCR amplification and subsequent sequence determination of the p24 gene, the relatedness of Borna disease virus (BDV) in various naturally infected animal species was determined. These results are indicative of a common ancestral virus pool and a remarkably low species barrier of BDV. Comparison of 11 sequences to that of tissue culture adapted virus revealed that the homology among all isolates was at least 96.2% at the nucleotide level, and 97% at the amino acid level. Viral sequences from sheep, donkey and horse were found to be not more distantly related to each ot...
Insulin-like growth factor II in the horse: determination of a cDNA nucleotide sequence and expression in fetal and adult tissue.
General and comparative endocrinology    November 1, 1994   Volume 96, Issue 2 270-275 doi: 10.1006/gcen.1994.1182
Otte K, Engström W.Horse cDNA for insulin-like growth factor II (IGF II) has been isolated. cDNA was synthesized from bulk mRNA and subsequently PCR-amplified and sequenced. Like its human counterpart, the mature horse IGF II peptide contains 67 amino acids with only two substitutions, isoleucine instead of valine in position 35 and asparagine instead of serine in position 36. The nucleotide homology was 92.1% between horse and human and 87.8% between horse and mouse. The isolated cDNA hybridized to multiple transcripts in fetal and adult tissues, thus confirming earlier reports on developmental expression of th...
GRASP: a novel heparin-binding serum glycoprotein that mediates oligodendrocyte-substratum adhesion.
Journal of neuroscience research    November 1, 1994   Volume 39, Issue 4 457-473 doi: 10.1002/jnr.490390413
Schirmer EC, Farooqui J, Polak PE, Szuchet S.Cell-substratum adhesion plays a crucial part in the cascade of events that control growth or turn on and consummate a differentiation program. We are investigating the molecular basis of oligodendrocyte (OLG) cytodifferentiation, employing pure cultures of OLGs isolated from postmyelination brains. We have shown that such OLGs will regenerate in vitro and reenact the ontogenic development of myelin, but to do so they need a signal. Adherence to a polylysine surface in the presence of 20% horse serum generates such a signal. Among the events that are turned on upon OLG adhesion is the phosphor...
Equilibrium unfolding studies of horse muscle acylphosphatase.
European journal of biochemistry    November 1, 1994   Volume 225, Issue 3 811-817 doi: 10.1111/j.1432-1033.1994.0811b.x
Taddei N, Buck M, Broadhurst RW, Stefani M, Ramponi G, Dobson CM.The stability and equilibrium unfolding behaviour of horse muscle acylphosphatase have been studied by denaturing the protein under various conditions of temperature, pH, and urea concentration. Far-ultraviolet circular dichroism (CD) and nuclear magnetic resonance (NMR) spectroscopy indicate that this small monomeric protein unfolds reversibly and cooperatively. Thermodynamic parameters, the Gibbs free energy delta G and enthalpy delta H of unfolding, have been estimated for denaturation of the protein from NMR and CD data as 19 kJ mol-1 and 350 kJ mol-1, respectively. CD and 1H-NMR results s...
Structural protein relationships among eastern equine encephalitis viruses.
The Journal of general virology    November 1, 1994   Volume 75 ( Pt 11) 2897-2909 doi: 10.1099/0022-1317-75-11-2897
Strizki JM, Repik PM.We have re-evaluated the relationships among the polypeptides of eastern equine encephalitis (EEE) viruses using SDS-PAGE and peptide mapping of individual virion proteins. Four to five distinct polypeptide bands were detected upon SDS-PAGE analysis of viruses: the E1, E2 and C proteins normally associated with alphavirus virions, as well as an additional more rapidly-migrating E2-associated protein and a high M(r) (HMW) protein. In contrast with previous findings by others, the electrophoretic profiles of the virion proteins of EEE viruses displayed a marked correlation with serotype. The pro...
The complete mitochondrial DNA sequence of the horse, Equus caballus: extensive heteroplasmy of the control region.
Gene    October 21, 1994   Volume 148, Issue 2 357-362 doi: 10.1016/0378-1119(94)90713-7
Xu X, Arnason U.The sequence of the mitochondrial (mt) DNA of the horse (Equus caballus) was determined. The length of the sequence presented is 16,660 bp. This figure, however, is not absolute due to pronounced heteroplasmy caused by variable numbers of the motif GTGCACCT in the control region of different molecules. Boundaries of the 13 peptide-coding genes were determined by the presence of start and stop codons, and by analogy with other eutherian mtDNAs. Three genes (COIII, NADH3 and NADH4) were not terminated by a stop codon. Comparison among the peptide-coding genes of the horse and eight other mammals...
Genetic Bit Analysis: a solid phase method for typing single nucleotide polymorphisms.
Nucleic acids research    October 11, 1994   Volume 22, Issue 20 4167-4175 doi: 10.1093/nar/22.20.4167
Nikiforov TT, Rendle RB, Goelet P, Rogers YH, Kotewicz ML, Anderson S, Trainor GL, Knapp MR.A new method for typing single nucleotide polymorphisms in DNA is described. In this method, specific fragments of genomic DNA containing the polymorphic site(s) are first amplified by the polymerase chain reaction (PCR) using one regular and one phosphorothioate-modified primer. The double-stranded PCR product is rendered single-stranded by treatment with the enzyme T7 gene 6 exonuclease, and captured onto individual wells of a 96 well polystyrene plate by hybridization to an immobilized oligonucleotide primer. This primer is designed to hybridize to the single-stranded target DNA immediately...
Monocyte maturation controls expression of equine infectious anemia virus.
Journal of virology    October 1, 1994   Volume 68, Issue 10 6270-6279 doi: 10.1128/JVI.68.10.6270-6279.1994
Maury W.In vivo, equine infectious anemia virus (EIAV) replicates in tissues rich in macrophages, and it is widely believed that the tissue macrophage is the principal, if not sole, cell within the host that replicates virus. No viral replication has been detected in circulating peripheral blood monocytes. However, proviral DNA can be detected in these cells, and monocytes may serve as a reservoir for the virus. In this study, an in vitro model was developed to clarify the role of monocyte maturation in regulating EIAV expression. Freshly isolated, nonadherent equine peripheral blood monocytes were in...
Characterization of a novel streptokinase produced by Streptococcus equisimilis of non-human origin.
Thrombosis and haemostasis    October 1, 1994   Volume 72, Issue 4 595-603 
Nowicki ST, Minning-Wenz D, Johnston KH, Lottenberg R.Streptokinases are proteins with plasminogen activator activity produced by certain hemolytic streptococci. We previously identified equine streptococcal isolates which produced streptokinases (ESKs) that bound both human and equine plasminogen but only readily activated equine plasminogen (14). This property was exploited to purify a representative ESK produced by Streptococcus equisimilis strain 87-542-W. Affinity chromatography with human plasminogen resulted in the isolation of a M(r) approximately 49,000 molecule with two isoforms. This ESK was subsequently compared to well characterized ...
Multiple genotypes of mitochondrial DNA within a horse population from a small region in Yunnan Province of China.
Biochemical genetics    October 1, 1994   Volume 32, Issue 9-10 371-378 doi: 10.1007/BF02426899
Wang W, Liu AH, Lin SY, Lan H, Su B, Xie DW, Shi LM.mtDNA genotypes of six domestic horses (three adult short horses whose heights are under 1 m and three common domestic horses) from a small region of 15 km2 in Malipo county of Yunnan province of China were investigated by the technique of restriction fragment length polymorphism (RFLP) with 16 restriction endonucleases which recognize 6-bp sequences. An average of 56 fragments for an individual was obtained. Unlike other domestic animals, this population of horses exhibits high mtDNA genetic diversity. Each of the six horses has a specific mtDNA genotype showing a pattern of multiple maternal...
Further characterisation of forms of haemosiderin in iron-overloaded tissues.
European journal of biochemistry    October 1, 1994   Volume 225, Issue 1 187-194 doi: 10.1111/j.1432-1033.1994.00187.x
Ward RJ, Ramsey M, Dickson DP, Hunt C, Douglas T, Mann S, Aquad F, Peters TJ, Crichton RR.The biochemical and biophysical properties of isolated haemosiderins have been compared to that of another iron-containing protein, termed prehaemosiderin, which sediments through chaotropic potassium iodide only after 20 h of ultracentrifugation, in contrast to that of haemosiderin which is recovered after 2 h of ultracentrifugation. The iron/protein ratio and iron/phosphate ratio were less that that of the corresponding haemosiderin, while the elemental composition was also reduced in many of the prehaemosiderin samples. Mossbauer spectroscopy and electron diffraction identified the predomin...
[Equine infectious arteritis: molecular biology, epidemiology and preventative measures].
Revue scientifique et technique (International Office of Epizootics)    September 1, 1994   Volume 13, Issue 3 845-854 
Zientara S.After a brief historical account of the outbreak of infectious arteritis of horses which occurred in 1984 in Kentucky (United States of America), the author reports on the present state of knowledge concerning the organisation of the genome of the virus. Clinical signs of the disease are described, as well as modes and routes of transmission. Finally, currently-available vaccination procedures are discussed and their value is assessed.
A new type of staphylococcal exfoliative toxin from a Staphylococcus aureus strain isolated from a horse with phlegmon.
Infection and immunity    September 1, 1994   Volume 62, Issue 9 3780-3785 doi: 10.1128/iai.62.9.3780-3785.1994
Sato H, Matsumori Y, Tanabe T, Saito H, Shimizu A, Kawano J.A new type of staphylococcal exfoliative toxin (sET) was isolated from the culture filtrate of a Staphylococcus aureus strain isolated from a horse with skin infection including phlegmon. The new sET was purified by precipitation with 80% saturated ammonium sulfate, column chromatography on DEAE-cellulofine A-500, gel filtration on a Sephadex G-75 column, and polyacrylamide gel electrophoresis (7.5% polyacrylamide). The new sET elicited general exfoliation of the epidermis with the so-called Nikolsky sign when inoculated into both 3-day-old mice and 1-day-old chicks, whereas sETA and sETB from...
Pathophysiology of sodium channelopathies: correlation of normal/mutant mRNA ratios with clinical phenotype in dominantly inherited periodic paralysis.
Human molecular genetics    September 1, 1994   Volume 3, Issue 9 1599-1603 doi: 10.1093/hmg/3.9.1599
Zhou J, Spier SJ, Beech J, Hoffman EP.It is often suggested that polygenic or environmental factors are responsible for clinical variability between patients with identical mutations. However, most dominant diseases are caused by a change-of-function alteration in the mutant allele's protein product. All patients are heterozygous and presumably express both mutant and normal proteins from the corresponding genes. Thus, a possible molecular mechanism for clinical variability could be the difference in relative levels of mutant vs. normal mRNA in different patients with the same mutation. To investigate this hypothesis, it is necess...
Proteolytic processing of the replicase ORF1a protein of equine arteritis virus.
Journal of virology    September 1, 1994   Volume 68, Issue 9 5755-5764 doi: 10.1128/JVI.68.9.5755-5764.1994
Snijder EJ, Wassenaar AL, Spaan WJ.To study the proteolytic processing of the equine arteritis virus (EAV) replicase open reading frame 1a (ORF1a) protein, specific antisera were raised in rabbits, with six synthetic peptides and a bacterial fusion protein as antigens. The processing of the EAV ORF1a product in infected cells was analyzed with Western blot (immunoblot) and immunoprecipitation techniques. Additional information was obtained from transient expression of ORF1a cDNA constructs. The 187-kDa ORF1a protein was found to be subject to at least five proteolytic cleavages. The processing scheme, which covers the entire OR...
Potency of partially purified malleo-proteins for mallein test in the diagnosis of glanders in equines.
Veterinary microbiology    August 15, 1994   Volume 41, Issue 4 391-397 doi: 10.1016/0378-1135(94)90035-3
Verma RD, Venkateswaran KS, Sharma JK, Agarwal GS.Malleo-proteins from synthetic broth mallein of six strains of Pseudomonas mallei were separated by trichloroacetic acid precipitation, ammonium sulfate precipitation and Ultrogel AcA 34 gel filtration chromatography. When tested comparatively with Dutch PPD mallein as standard on P. mallei-sensitized and normal horses all the strains were found to be malleinogenic, trichloroacetic acid precipitated proteins were comparable to Dutch PPD mallein in potency and innocuity whereas ammonium sulfate-precipitated proteins elicited non-specific reactions. Ultrogel AcA 34 chromatographed high molecular...
Molecular cloning and expression of two horse pancreatic cDNA encoding colipase A and B.
Biochimica et biophysica acta    August 4, 1994   Volume 1213, Issue 3 357-360 doi: 10.1016/0005-2760(94)00096-4
Crenon I, Granon S, Chapus C, Kerfelec B.Pancreatic colipase plays an essential role in the intestinal fat digestion by anchoring lipase on lipid/water interfaces in the presence of bile salts. In contrast to other species, two molecular forms of colipase, A and B, have been found in horse. The two corresponding cDNAs were isolated from a horse pancreatic library and their nucleotide sequences were determined. Moreover, for the first time, active colipase has been obtained after transfection of COS cells by either colipase A or B cDNA.
Detection of aberrant alleles in the D-loop region of equine mitochondrial DNA by single-strand conformation polymorphism (SSCP) analysis.
Animal genetics    August 1, 1994   Volume 25, Issue 4 287 doi: 10.1111/j.1365-2052.1994.tb00211.x
Ishida N, Hirano T, Mukoyama H.No abstract available
Equine dinucleotide repeat polymorphism at the VIAS-H7 locus.
Animal genetics    August 1, 1994   Volume 25, Issue 4 292 doi: 10.1111/j.1365-2052.1994.tb00221.x
Ewen KR, Matthews ME.No abstract available
Application of PCR to a clinical and environmental investigation of a case of equine botulism.
Journal of clinical microbiology    August 1, 1994   Volume 32, Issue 8 1986-1991 doi: 10.1128/jcm.32.8.1986-1991.1994
Szabo EA, Pemberton JM, Gibson AM, Thomas RJ, Pascoe RR, Desmarchelier PM.PCR for the detection of botulinum neurotoxin gene types A to E was used in the investigation of a case of equine botulism. Samples from a foal diagnosed with toxicoinfectious botulism in 1985 were reanalyzed by PCR and the mouse bioassay in conjunction with an environmental survey. Neurotoxin B was detected by mouse bioassay in culture enrichments of serum, spleen, feces, and intestinal contents. PCR results compared well with mouse bioassay results, detecting type B neurotoxin genes in these samples and also in a liver sample. Other neurotoxin types were not detected by either test. Clostrid...
The aspartic proteinase of equine infectious anaemia virus.
Biochemical Society transactions    August 1, 1994   Volume 22, Issue 3 304S doi: 10.1042/bst022304s
Powell DJ, Le Grice SJ, Kay J.No abstract available
[Morphologic and molecular biologic studies of the etiology of equine sarcoid].
Tierarztliche Praxis    August 1, 1994   Volume 22, Issue 4 368-376 
Teifke JP.From 932 equine skin lesions 421 were diagnosed as sarcoids (about 45%). The most common locations were the ventral body regions, head, neck and sites of thin skin. Most often the fibroblastic type, less frequently the mixed type and most infrequent the verrucous type of sarcoid were diagnosed. Detection of BPV-DNA was performed by polymerase chain reaction (PCR) using an oligonucleotide primer pair located in the E5-open reading frame. DNA of BPV 1 and BPV 2 could be differentiated by digestion with restriction endonucleases. In 97 out of 108 sarcoids BPV-DNA was detected by PCR. Most samples...
Molecular weight alterations of alpha-1 proteinase inhibitor in equine bronchoalveolar lavage fluid.
Electrophoresis    August 1, 1994   Volume 15, Issue 8-9 1193-1197 doi: 10.1002/elps.11501501180
Milne EM, Pemberton AD, McGorum BC, Dixon PM, Miller HR.The equine alpha-1 proteinase inhibitor (alpha 1PI) system differs from that of man in that the equine system consists of four closely-linked genes (Spi1-Spi4) whereas in man, a single gene encodes for alpha 1PI. We have previously found differences in the proportion of the Spi proteins in equine serum and bronchoalveolar lavage fluid (BALF). We therefore wished to determine whether, as reported in man, there was any molecular weight difference between the Spi proteins in serum and BALF. alpha 1PI and albumin from equine BALF migrated further towards the anode compared with serum alpha 1PI on ...
Polymorphic sequence in the D-loop region of equine mitochondrial DNA.
Animal genetics    August 1, 1994   Volume 25, Issue 4 215-221 doi: 10.1111/j.1365-2052.1994.tb00196.x
Ishida N, Hasegawa T, Takeda K, Sakagami M, Onishi A, Inumaru S, Komatsu M, Mukoyama H.The D-loop regions in equine mitochondrial DNA were cloned from three thoroughbred horses by polymerase chain reaction (PCR). The total number of bases in the D-loop region were 1114 bp, 1115 bp and 1146 bp. The equine D-loop region is A/T rich like many other mammalian D-loops. The large central conserved sequence block and small conserved sequence blocks 1, 2 and 3, that are common to other mammals, were observed. Between conserved sequence blocks 1 and 2 there were tandem repeats of an 8 bp equine-specific sequence TGTGCACC, and the number of tandem repeats differed among individual horses....
Pregnancy-associated endometrial expression of antileukoproteinase gene is correlated with epitheliochorial placentation.
Molecular reproduction and development    August 1, 1994   Volume 38, Issue 4 357-363 doi: 10.1002/mrd.1080380402
Badinga L, Michel FJ, Fields MJ, Sharp DC, Simmen RC.Uterine expression of the mRNA encoding antileukoproteinase (ALP) is highest in pig uterus during mid- to late pregnancy, suggesting a stage of pregnancy-dependent role for this elastase/cathepsin G protease inhibitor in feto-maternal interactions. To examine a potential relationship between uterine synthesis of ALP and the type of placentation in mammalian species, the expression of ALP mRNA and/or protein in pregnant mares, cows, rats, and mice was evaluated. Genomic DNA and mRNA hybridization analyses were performed using a porcine ALP cDNA as probe. The concentration of ALP protein in repr...
Light chain isotype regulation in the horse. Characterization of Ig kappa genes.
Journal of immunology (Baltimore, Md. : 1950)    August 1, 1994   Volume 153, Issue 3 1099-1111 
Ford JE, Home WA, Gibson DM.Horse Ig kappa genes have been characterized to determine whether there may be a structural basis for the low level of kappa expression in this species. The overall organization of the J kappa-C kappa locus is remarkably similar to that of the mouse and human loci. A single C kappa exon is separated by 2.9 kb from five J kappa segments, four of which seem functional and three of which are associated with canonical recombination signal sequences. A highly conserved intron enhancer was identified upstream of the C kappa exon and a single restriction fragment in horse genomic DNA hybridized stron...
Molecular dynamics simulation of equine infectious anemia virus Tat protein in water and in 40% trifluoroethanol.
Journal of biomolecular structure & dynamics    August 1, 1994   Volume 12, Issue 1 019-36 
Sticht H, Willbold D, Rösch P.Two molecular dynamics (MD) simulations were performed in order to increase the understanding of the dependence of protein conformation on solvent environment. The protein used for these simulations is the transcriptional activator of the equine infectious anemia virus (EIAV-Tat). The structure of this protein has been determined by nuclear magnetic resonance (NMR) in aqueous solution (Willbold et al., Science 264, 1584 (1994)) and in 40% (v/v) trifluoroethanol (TFE) (Sticht et al., Eur. J. Biochem., submitted) showing considerable differences in the stability of the secondary structure elemen...
Characterization of monoclonal antibodies specific for equine homologues of CD3 and CD5.
Immunology    August 1, 1994   Volume 82, Issue 4 548-554 
Blanchard-Channell M, Moore PF, Stott JL.Two monoclonal antibodies (mAb), UC F6G-3 and UC F13C-5, were characterized as being specific for the apparent equine homologues of CD3 and CD5, respectively. Both antibodies exhibited characteristics of pan-T-lymphocyte markers based upon immunohistology and two-colour flow cytometry. UC F6G-3 precipitated a complex of proteins (up to seven) with molecular weights ranging from 18,000 to 42,000, similar to the human and murine CD3 complex. Upon further dissociation of the precipitated complex, two proteins were identified with molecular weights of 22,000 and 27,000. Immobilized UC F6G-3 was ef...
Models of the three-dimensional structures of echidna, horse, and pigeon lysozymes: calcium-binding lysozymes and their relationship with alpha-lactalbumins.
Journal of protein chemistry    August 1, 1994   Volume 13, Issue 6 569-584 doi: 10.1007/BF01901539
Acharya KR, Stuart DI, Phillips DC, McKenzie HA, Teahan CG.Similarities in amino acid sequences, three-dimensional structures, and the exon-intron patterns of their genes have indicated that c-type lysozymes and alpha-lactalbumins are homologous proteins, i.e., descended by divergent evolution from a common ancestor. Like the alpha-lactalbumins, echidna milk, horse milk, and pigeon eggwhite lysozymes all bind Ca(II). Models of their three-dimensional structures, based on their amino acid sequences and the known crystal structures of domestic hen eggwhite and human lysozymes and baboon and human alpha-lactalbumins, have been built. The several structur...