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We have cloned and sequenced the gene encoding porcine E-selectin. The gene comprises 12 exons and 11 introns. Two pseudoexons are contained within intron 4 and intron 6. These sequences are similar to the corresponding exons in the human E-selectin sequence; however, they are not present in the porcine E-selectin-encoding cDNA. Transcription starts at position -498 relative to the translation...
A synthetic oligodeoxynucleotide probe based on a highly conserved region of the sigma factors was used to identify and clone the rpoD gene encoding the principal sigma factor of R. capsulatus. The deduced polypeptide contains 674 amino acids and has a predicted molecular mass of 75 942 Da. The deduced amino acid sequence of R. capsulatus RpoD protein exhibits 46.2% and 45.7% identity with housekeeping...
A murine genomic clone encoding the relA (p65 NF-κB) locus was isolated and characterized. The clone spanned about 20 kb and included at least 10 exons and the relA promoter. Sequencing of the promoter region revealed the presence of a murine-specific intron of 546 bp intervening the homologue of human exon 1. The species-specific organisation of relA genes and promoter elements is compared and...
The nucleotide sequence of a 3.1-kb genomic DNA fragment carrying the omp3, omp2 and srp gene homologs from Chlamydia psittaci strain GPIC was determined. A comparative analysis of the GPIC sequence with other chlamydial omp2-linked sequences reveals highly conserved omp3 and omp2 upstream sequences across species, suggesting a unified mechanism of transcription regulation. In contrast, the omp2-srp...
The nucleotide (nt) sequence encoding the bovine homologue of interleukin-5 (boIL-5) was determined. Total cDNA generated from stimulated bovine peripheral blood lymphocytes was used to amplify the boIL-5 gene using primers based on the 5 and 3 untranslated regions of the ovine IL-5 cDNA sequence. The boIL-5 coding sequence is 405 bp long, coding for a 15.2 kDa precursor protein of 134 amino...
The secA gene of Streptomyces lividans was cloned using as probe a 57-mer oligonucleotide based on conserved sequences of the Escherichia coli secA and the Bacillus subtilis div genes. It encodes a protein of 946 amino acids (aa) with a deducedM r of 106 079, with high similarity to all known SecA proteins. All the previously described conserved motifs of SecA proteins were conserved in...
A repetitive HindIII fragment of DNA from Raja montagui (Rajiformes) was cloned and sequenced for the first time in cartilaginous fishes. This element, which comprises approximately 5% of the whole genome of the spotted ray, is absent in long tandem arrays, being typical of satellite DNA. It appeared constituted by 311 AT-rich bp (61%). The clone was hybridized to the genomic DNA of species with...
The pSUM vectors, a family of Mycobacterium/Escherichia coli shuttle cloning vectors derived from the pSU series of vectors are described. They all contain the pACYC184 origin of replication enabling them to replicate in E. coli, the mycobacterial pAL5000 origin of replication and a kanamycin-resistance (Km R ) gene which allows selection in both E. coli and mycobacteria. They carry a multiple...
Analysis of replication intermediates produced by in vitro replication of the Bacillus subtilis oriC plasmid revealed that replication initiated at an untranslatable DnaA box region downstream of the dnaA gene. In order to show that replication of the B. subtilis chromosome also starts at the same region in vivo, we have analyzed replication intermediates generated in vivo by the two-dimensional...
With use of the signal sequence trap method, we isolated a cDNA encoding a novel secretory protein, SDF-2, from the mouse stromal cell line, ST2. The human homologue of SDF-2 was also isolated. The amino acid (aa) sequences deduced from both the clones were conserved more than 92%. The chromosomal localization of the human SDF-2 gene was mapped to 17q11.2. The aa sequence of SDF-2 shows similarity...
The nucleotide sequence of nearly 6 kb of genomic DNA located immediately upstream of the omp3-omp2 operon of Chlamydia psittaci strain GPIC was obtained, revealing four significant open reading frames (ORFs), named ORF1, ORF2, ORF4 and ORF5. Searches for homologous sequences in the GenBank/EMBL databases have revealed that: (a) the open-ended ORF1 putatively encodes an homolog of RecJ of Escherichia...
Introduction of in vitro transcribed mRNA into Xenopus laevis embryos is a useful technique for analyzing gene function. In order to optimize expression of cDNA constructs from in vitro transcribed mRNAs, we examined the translational efficiency of reporter genes that simulated cDNAs synthesized by tailing with deoxyguanosine (dG) before second strand cDNA synthesis. When transcribed in vitro, these...
The SSD1 gene of Saccharomyces (S.) cerevisiae is a polymorphic gene involved in many aspects of the yeast cell growth (Sutton et al., 1991). We found that ssd1 null mutant shows increased sensitivities of growth to trifluoperazine (TFP) and high concentration of Ca 2+ . A high-copy suppressor gene, HST1, for the TFP and Ca 2+ sensitivities of ssd1 null mutant was cloned...
We have investigated the requirements necessary for high-level production of the hepatitis B virus (HBV strain ayw) large surface glycoprotein preS1 with vaccinia virus (VV) recombinants. In earlier studies, only nanogram amounts of preS1 could be obtained from cells infected with an appropriate recombinant VV carrying the preS1 gene under the transcriptional control of a conventional VV promoter...
We have previously identified the α element within the mouse H2A and H3 histone gene coding region activating sequences (CRAS). This common element is required for normal in vivo expression of these two replication-dependent genes and interacts with nuclear factor(s). Here we report that the CRAS α element is present in the coding region sequences of two other replication-dependent mouse H genes,...
Anti-Mullerian hormone (AMH) is responsible for regression of the Mullerian ducts in males during embryonic development. This peptide hormone of the transforming growth factor-β family is also believed to play a broader role in sex determination, affecting differentiation and morphogenesis of the testes. Accordingly, in mammals, AMH is produced at much higher levels in male fetuses than in female...
Metallothionein (MT) cDNAs were cloned and sequenced from two genera of ducks, Muscovy (Cairina muschata) and Tsai ya (Anas platyrhynchos). The two cDNAs show an extremely high sequence homology and contain an open reading frame encoding 63 amino acids. MT mRNA expressions were studied after metal induction using the cloned cDNA as a probe. Cadmium and copper induced MT gene efficiently, whereas...
The lysis genes of a Lactobacillus phage φg1e were cloned, sequenced, and expressed in Escherichia coli. Nucleotide sequencing of a 3813-bp φg1e DNA revealed five successive open reading frames (ORF), Rorf50, Rorf118, hol, and lys andRorf175 , in the same DNA strand. By comparative analysis of the DNA sequence, the putative hol product (holin) has an estimated molecular weight is 14.2 kDa, and contains...
A number of unrelated protein-encoding genes from sulfothermophilic archaea, Sulfolobus acidocaldarius, Sulfolobus solfataricus, Pyrococcus furiosus and Pyrococcus woesei, has been analyzed. In the Sulfolobus genus, the content of A+T is significantly higher than that of C+G and the base usage follows the order, A>T>G>C. In Pyrococcus, the A+T content is also higher than that of C+G, but...
By utilizing the oxygen-sensitive Escherichia coli Mn-superoxide dismutase (Mn-SOD) promoter, we have developed a vector system that expresses high levels of cloned foreign genes. The promoter for the bacterial Mn-SOD, as well as both 5 -untranslated and transcriptional termination sequences were ligated to a synthetic linker containing two restriction enzyme cloning sites. The vector also contained...
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