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The MIP (major intrinsic protein) gene, a member of an ancient family of membrane channel genes, encodes the predominant fiber cell membrane protein of the ocular lens. Its specific expression in the lens fibers is temporally and spatially regulated during development. To study the regulation of expression of MIP and delineate the regulatory elements underlying its tissue specificity and ontogenic...
The argG gene of Streptomyces clavuligerus (Scl) has been cloned by complementation of argG mutants ofEscherichiacoli and S. lividans (Sl). The argG nucleotide (nt) sequence showed that it corresponds to a new type ofargG different from the corresponding genes of S. coelicolor (Sco) and Sl. It encodes a 43 250-Da protein that showed higher similarity to argininosuccinate synthetases (ASS) from Methanococcus...
Glucagon was found to increase the mRNA level of the uricase-encoding gene (UOX), but not that of genes encoding other peroxisomal enzymes, such as catalase, acyl-CoA oxidase and enoyl-CoA hydratase/3-hydroxyacyl-CoA dehydrogenase. The possible involvement of cAMP in the glucagon-induced transcription of rat UOX was studied by measuring the enhancer activity of the isolated 5 -untranslated region...
A novel expression vector (pAVEX16C) has been constructed that directs the synthesis of desired polypetides as fusions with the C terminus of chicken egg-white avidin (Avd). With this and a commercial GST gene (encoding glutathione S-transferase) fusion vector (pGEX-3X, Pharmacia), we produced Avd as fusions C- and N-terminally linked to GST in Escherichia coli. By using the Avd tail and a simple...
The nucleotide (nt) sequence of a 9704-bp EcoRI fragment of Streptococcus pneumoniae (Sp) type-3 DNA has been determined and found to contain one partial and five complete open reading frames (ORFs). One of these ORFs corresponds to the cap3A gene coding for the UDP-glucose (UDPGlc) dehydrogenase which is directly responsible for the transformation of some unencapsulated serotype-3 Sp mutants to...
We have investigated actions of the Wilms' tumour suppressor zinc finger transcription factor, WT1, on promoters of the mouse insulin-like growth factor II-encoding gene (Igf-2). Two variant forms of WT1 repressed the two majorIgf-2 promoters (P2 and P3) in transient transfection assays. WT1-binding sites were characterised in both these promoters and in the transcribed region downstream from P2,...
We have identified a 236-bp first intron segment of the mouse adenosine deaminase gene (ADA) that shares 71.1% identity with the human ADA thymic enhancer. This segment has the same natural orientation as the human enhancer and a relative location within the first intron very analogous to that of the human enhancer. Four highly conserved regions were defined within this segment, including a 72-bp...
Avian cyclin D2 (Cyl D2)-encoding cDNA clones were isolated from a chicken UG9 T-cell λgt10 library. Sequence analysis revealed a high degree of sequence conservation with both the mouse and human Cyl D2, and somewhat lower similarity with the mouse and human Cyl D1 and D3. The homology is highest between species in the Cyl-box domain which is well conserved among human, mouse and chicken. A single...
Phage display of proteins can be used to study ligand-receptor interaction and for the affinity-maturation of binding sites in polypeptide hormones and/or cytokines. We have expressed human interleukin-6 (hIL-6) on M13 phage in a monovalent fashion as a fusion protein with the phage coat protein, pIII. Phage-displayed hIL-6 is correctly folded, as judged by its ability to interact with conformation-specific...
By using a P22 phage-mediated cloning system, the nrdAB genes of Salmonella typhimurium (St), encoding a ribonucleotide reductase (RR) of class I, have been isolated. The coding regions of the St nrdAB operon show a very high identity with those of the homologous operon of Escherichia coli (Ec). Nevertheless, there are significant differences in their promoter regions since, although the promoters...
An 1.7-kb Arabidopsis thaliana (At) cDNA was isolated by complementation of a bap1 mutation affecting the transport of branched-chain amino acids (aa) in the yeast Saccharomyces cerevisiae. The determination of the nucleotide (nt) sequence revealed an open reading frame of 1383 nt which may encode a protein of 461 aa with a predicted molecular mass of 51 038 Da. The deduced aa sequence exhibited...
Two genes that encode methylmalonyl-CoA mutase (MCM) have been characterised in Porphyromonas gingivalis W50 (Pg). The genes, designated mcmA and mcmB are transcribed in an operon and encode the MCM small subunit (SS, 68 626 Da) and the MCM large subunit (LS, 78 703 Da), respectively. A recombinant Escherichia coli (Ec) clone harbouring thePg mcmA and mcmB genes expressed MCM activity 280-times...
Previous studies have shown that 13 genes located in a 14.6-kb region of the chromosome of Staphylococcus aureus (Sa) M are required for type-1 capsular polysaccharide (CP1) biosynthesis. In this report, a total of 17 Sa strains producing different CP serotypes were analyzed by Southern hybridization using DNA probes from the cap1 coding region and the flanking sequences. The results showed that...
The gene (γFBG) encoding the fibrinogen γ chain (γFBG) has been shown to exhibit both tissue-specific and ubiquitous expression. To confirm the identity of the γFBG transcripts expressed in extrahepatic tissue, lung tissue was chosen as a model of extrahepatic γFBG gene expression. A ferret lung cDNA clone bank was constructed in λgt11 and several positive plaques were isolated using cross-species...
We report the isolation of a cDNA encoding a new type of transcription factor S-II, termed h-SII-T1, from a human library. The mRNA corresponding to the clone is highly expressed in testis and ovary. Comparison of the deduced amino acid (aa) sequence with those of other S-II molecules shows that (i) the C-terminal zinc finger (Zf) domain is highly conserved, and (ii) the central segment is most...
A 40-kDa subunit in eukaryotic RNA polymerases (Pol) I and III (e.g., yeast yAC40) is related in a part of its aa sequence to the α subunit of prokaryotic Pol and to a 35-44-kDa subunit in Pol II (e.g., yeast yB44). We have cloned two cDNAs, AtRPAC42 and AtRPAC43, from an Arabidopsis thaliana (At) (ecotype Columbia) λ Yes expression library that encode Pol I and III subunits related to yAC40. The...
Adducin (ADD) is a heterodimeric protein of the membrane skeleton with subunits of 103 (α) and 97 kDa (β). It promotes the assembly of the spectrin-actin network. We have previously shown that one point mutation in each of the α and β rat ADD-encoding genes is associated with blood pressure variation in an animal model for hypertension, the Milan hypertensive strain of rats, probably due to a change...
The nucleotide sequence of the Borrelia burgdorferi (Bb) Sh-2-82 recA gene has been determined using PCR-based approaches without the construction of a genomic library. The gene should encode a protein of 365 amino acids which is highly homologous to other known RecA proteins. It represents a new homolog from a distinct phylogenetic branch of eubacteria. Although, previous reports concluded that...
The alcA gene, essential for the production of the dihydroxamate siderophore, alcaligin, by Bordetella bronchiseptica, was cloned and sequenced. The alcA gene was identified on a 4.7-kb EcoRI genomic fragment adjacent to a Tn5lac transposon insertion that inactivated alcaligin production in strain MBORD846. Analysis of the alcA nucleotide sequence revealed a putative Fur-binding site, suggesting...
We report the isolation and characterization of a gene (designated mcp60) encoding the mitochondrial (mt) 60-kDa heat-shock protein (HSP60) in the fission yeast Schizosaccharomyces pombe. The deduced amino-acid sequence (582 aa) of this gene is highly similar to the known mt HSP60 from diverse organisms. When its sequence was related to the known functional domains of bacterial HSP60 (GroEL), the...
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