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The green fluorescent protein (GFP) from the jellyfish, Aequorea victoria, has become a versatile reporter for monitoring gene expression and protein localization in a variety of cells and organisms. GFP emits bright green light (λ max = 510 nm) when excited with ultraviolet (UV) or blue light (λ max = 395 nm, minor peak at 470 nm). The chromophore in GFP is intrinsic...
Hepatocyte nuclear factor 4 (HNF-4) is an essential positive regulator of a large number of liver-specific genes. We report here the isolation of three HNF-4 isoforms from a human liver cDNA library. hHNF-4A and hHNF-4B, differing by the insertion of 10 amino acids in the C-terminal region, have been previously identified in mouse, rat and human liver. The novel isoform, hHNF-4C, is identical to...
Mitochondrial aldehyde dehydrogenase (ALDH2) activity is produced at low levels in many tissues, with highest production in liver. Transfection assays using the first 600 bp of upstream DNA provided evidence for both positive and negative regulatory elements in the proximal promoter. A region from - 79 to - 116 bp was protected in DNase I footprinting assays and bound in electrophoretic mobility...
The H1 class of histones is implicated in organizing a higher order of chromatin structure and in involvement with transcriptional repression. They comprise multiple subtypes that presumably have a specific function. We report here the isolation and characterization of a human cDNA encoding a novel subtype of histone H1. It is the most distantly related subtype of mammalian H1 reported to date. ...
Retinoic acid (RA) plays a critical role in normal development, growth and differentiation by modulating the expression of target genes. Using substractive hybridization cloning, we isolated two cDNAs, whose corresponding mRNAs are repressed upon RA treatment of P19 embryonal carcinoma (EC) cells. The cDNAs correspond to the serine hydroxymethyltransferase (shmt) gene and the early transposon, ETnMG1...
Phenol sulfotransferases (PST) esterify hydroxylated aromatic compounds with sulfate, and therefore play a role in the metabolism of xenobiotics. In this investigation, a bovine PST gene of 2372 bp was amplified from genomic DNA. Sequence overlap with the cognate cDNA revealed seven exons, with all introns containing GT/AG splicing donor/acceptor sites. This product was subcloned into an SV40-based...
Transcription of genes from phage LL-H can be divided into an early phase and a late phase. The early gene region was located in a 5.9-kb segment of the phage LL-H genome and it was part of the sequence that completed the phage LL-H genome sequence, 34 659 bp in size. Phage LL-H is the first completely sequenced Lactobacillus phage. In the main coding strand of phage LL-H genome 48 putative ORFs...
Two protein kinase (PK) genes, gPK1 and gPK2, were cloned from the genome of the ancient protozoan parasite, Giardia duodenalis (Gd). Both gPK genes and their products are highly homologous (85% and 77% identical, respectively). gPK1 and gPK2 contain all the motifs characteristic of PK, but they are not highly homologous to other PK and therefore belong to a novel PK gene family. Northern blot...
The SPO1 gene of Saccharomyces cerevisiae has been cloned and sequenced. The Spo1 protein reveals significant similarity with fungal phospholipase B (PLB) enzymes. Features of the SPO1 gene sequence are presented.
Genes encoding the type I restriction-modification (R-M) system of the bovine pathogen, Pasteurella haemolytica, have been identified immediately downstream of a locus that encodes a transcriptional activator of P. haemolytica leukotoxin expression. Type I enzymes are encoded by three genes called hsdM, hsdS and hsdR, and have fallen into three groups, called Ia, Ib and Ic. HsdS provides a sequence...
We report the cDNA sequence encoding Ub52 from the cnidarian Acropora millepora. As in other eukaryotes, the N-terminal region of the deduced amino-acid sequence is a ubiquitin moiety and the C-terminal region CEP52, a protein component of the large ribosomal subunit. A. millepora Ub52 is highly homologous (>95% identical) with the corresponding Drosophila and vertebrate sequences, the cnidarian...
The in vitro amplification method for heterologous gene expression in mammalian cells is based on the stable transfection of cells with long, linear DNA molecules having several copies of complete expression units, coding for the gene of interest, linked to one terminal unit, coding for the selectable marker. DNA concatenamers containing additional expression units can also be prepared: we exploited...
The major capsid protein L1 of papillomaviruses expressed recombinantly or in infected cells has the intrinsic ability to form virus-like particles (VLPs) which display conformational epitopes necessary to elicit high-titered, virus-neutralizing antibodies. We have shown previously that the L1 gene of human papillomavirus type 6a (HPV6) can be expressed efficiently in Saccharomyces cerevisae (Sc)...
In this study, we describe the cloning of the mouse homologue of the orphan receptor, RZR/RORγ, a member of the nuclear receptor superfamily, from a mouse muscle cDNA library. The amino acid sequence of mouse RORγ (mRORγ) is highly homologous to that of human RORγ, with an overall identity of 88%. Northern blot analysis using RNA from different tissues showed that mRORγ was found to be highly expressed...
In Saccharomyces cerevisiae, the CDC2 gene encodes the large subunit of DNA polymerase III, the analogue of mammalian DNA polymerase δ. We have isolated DNA fragments from a library of Candida albicans genomic DNA in the vector pRS316 that rescue temperature sensitive cdc2 mutations in S. cerevisiae. These fragments contain an ORF coding for a protein of 1038 aa with a predicted molecular mass of...
Three PCR-amplified DNA fragments hybridizing with the OLE1 gene encoding Δ9-fatty acid desaturase of Saccharomyces cerevisiae were obtained using, respectively, genomic DNAs of one strain each of Kluyveromyces thermotolerans, Pichia angusta and Yarrowia lipolytica as templates. A gene designated P-OLE1 was cloned from the above fragment of P. angusta and sequenced. An open reading frame of P-OLE1...
Lens preferred-expression of the mouse αA-crystallin gene (αA-cry) is regulated at the transcriptional level by multiple elements located in the 5′ flanking region of the gene. Here we present the first analysis of the functional role of the mouse αA-cry +1 region and the protein(s) which bind to it. The −7/+5 region of this promoter exhibits sequence similarity with the consensus upstream stimulating...
A cDNA clone encoding a polypeptide with homology to the novel SPF1 DNA-binding protein of sweet potato has been isolated from a cDNA library from RNA of senescing cucumber (Cucumis sativus, L.) cotyledons. Comparison of the two sequences reveals similar features which may be important in the evolution and function of this protein, including a duplicated region of about 56 amino acids (aa). The first...
Tyrosine phosphorylation of proteins plays an important role in cellular signaling and many cellular activities. The levels of cellular phosphorylation are reversibly controlled by protein tyrosine kinases and protein tyrosine phosphatases. The murine R-PTP-κ, a receptor-type protein tyrosine phosphatase, has recently been cloned (Jiang et al. (1993)Mol. Cell. Biol. 13, 2942–2951). In order to identify...
New cloning and expression vectors that replicate both in Pasteurella haemolytica and in Escherichia coli were constructed based on a native sulfonamide (Su R ) and streptomycin (Sm R ) resistant plasmid of P. haemolytica called pYFC1. Each shuttle vector includes an MCS and a selectable antibiotic resistance marker that is expressed in both organisms. Plasmid pNF2176 carries theP...
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