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cDNA cloning, genomic structure and expression analysis of the bovine lanosterol 14alpha-demethylase (CYP51) in gonads.

Meiosis activating sterol (MAS), the intermediate of cholesterol biosynthesis, is an important substance to stimulate oocytes maturation in FSH-induced signal transduction pathway. Lanosterol 14alpha-demethylase (CYP51) converts lanosterol to MAS. Although MAS is firstly isolated from bovine testis, the information about bovine CYP51 gene and its expression is little. In present studies, the cDNA cloning, genomic structure, chromosomal mapping, and expression patterns of bovine CYP51 were demonstrated. The cDNA coding bovine CYP51 contains a 1509 bp open reading frame and a 1119 bp 3' untranslated region. And the bovine CYP51 gene includes 10 exons and spans about 17 kb. Screening the cattle RH5000 panel bovine CYP51 is mapped to chromosome 4 (0cR). The sequenced promoter region is TATA-less and contains several highly conserved regulatory elements, such as GC-box, cAMP-responsive elements (CRE), sterol regulatory element (SRE) which is important fragment for its transcription. No evidence of processed pseudogenes is found using long PCR and Southern blot. Northern blot analysis reveals that an approximately 2.7 kb mRNA is expressed in all the examined bovine tissues, while a 1.8 kb mRNA is found only in the mature bovine testis where the MAS is accumulated. Immunochemistry analysis shows that leydig cells express the highest level of the CYP51 protein in testis. Among different stages follicles it is localized primarily to the oocytes with the level varying slightly. Granulosa cells of primordial, primary and secondary follicles show background staining. While granulosa cells facing the antrum and cumulus granulosa cells of antral follicles show considerably heavier staining. The highest level is expressed in corpus lutea. These data indicate a stage- and cell type-specific expression of CYP51 protein in bovine oogenesis.

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Myristoyl-CoA:protein N-myristoyltransferases: isoform identification and gene expression in retina.

The mammalian myristoyl-CoA:protein N-myristoyltransferase (NMT) family consists of Type I and Type II enzymes that typically catalyze the addition of myristic acid (14:0) to the N-terminus of specific proteins using myristoyl-Coenzyme A as a donor. However, the N-terminus of certain proteins in frog and bovine retina are modified with fatty acids other than myristic acid, and this utilization of alternative acyl-CoAs has not been found in any other tissues. This alternative use of acyl groups is known as retina heterogeneous acylation and occurs by an unknown mechanism. Therefore, the focus of this work has been to identify NMT isoforms that may be unique or differentially expressed in bovine retina that may utilize acyl-CoAs in addition to myristoyl-CoA.

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DNA (cytosine 5) methyltr MIC2 Gene Protein, CD99; MIC2 Gene Protein, CD99; MIC2 Gene Protein, CD99; Acyl CoA binding Protein HIV 1 intergase antigen. T4 SSB (gene 32) protein T4 SSB (gene 32) protein T4 SSB (gene 32) protein T4 SSB (gene 32) protein Human Macrophage Inflamma Human Macrophage Inflamma

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Cytokine transcription in lymph nodes of cattle in different stages of bovine leukemia virus infection.

Bovine leukemia virus (BLV) is a transforming oncovirus that contains no oncogenes or preferred site of proviral integration. The role of cytokines in the disease process of BLV is potentially important due to the similarity of BLV with other retroviruses in which cytokines play a role, such as HTLV-I and -II. Mesenteric and supra-mammary lymph nodes were obtained from a panel of nine cattle. Three were non-infected controls, three were BLV-positive aleukemic (AL), and three were BLV-positive persistent lymphocytotic (PL). Mononuclear cells were perfused from the organs and total RNA extracted from either 1 x 10(8) unseparated cells or 1 x 10(7) purified CD4/CD8 T-cells. cDNA was generated and subjected to RT-PCR to analyze cytokine transcription during disease progression. cDNA levels were normalized using beta-actin PCR at sub-plateau cycle number, enabling a semi-quantitative assessment of cytokine gene transcripts. Using this approach, IL-2, IL-10 and IFN-gamma message was detected in the T-cell fractions of all of the BLV-infected animals, but not in the non-infected controls.

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Sequence, molecular properties, and chromosomal mapping of mouse lumican.

Lumican is a major proteoglycan of vertebrate cornea. This study characterizes mouse lumican, its molecular form, cDNA sequence, and chromosomal localization.

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Molecular cloning and chromosomal localization of human 4-beta-galactosyltransferase.

A cDNA clone to human 4-beta-galactosyltransferase (EC 2.4.1.38) was isolated from a human liver lambda gt11 expression library by using a monospecific polyclonal antiserum to affinity-purified bovine enzyme. The authenticity of this cDNA clone has been demonstrated by several criteria. Under conditions of chronic treatment with the beta-adrenergic receptor agonist isoproterenol, rat parotid glands show an approximately 10-fold increase in 4-beta-galactosyltransferase activity. The increased enzyme activity was reflected in dot-blot analysis of control and isoproterenol-treated rat parotid RNA by using the human cDNA as probe. Hybrid-selection and in vitro translation identified a protein product with a molecular mass of 47 kDa that was immunoprecipitated with the bovine antiserum. The full-length human cDNA clone was then isolated and the DNA sequence for the NH2-terminal portion of the protein was deduced. Comparison of the NH2-terminal protein sequence from the bovine protein with that of the human cDNA clone confirmed its identity. In addition, the human cDNA clone was used to localize the gene for 4-beta-galactosyltransferase to human chromosome 4 by Southern analysis of a somatic cell hybrid panel.

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