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In order to identify important regions of the human CAML (calcium-modulating cyclophilin ligand) protein, a cDNA encoding the chicken homologue was isolated using low-stringency hybridization screening of a T-lymphocyte cDNA library. The deduced amino acid sequences of chicken and human CAML show areas of extensive conservation, suggesting an evolutionarily important role in signaling.
A DNA segment from Mycobacterium tuberculosis containing a gene for a putative sigma factor was isolated and sequenced. The protein encoded by this gene is 92% similar to the Mycobacterium smegmatis sigma factor MysB, and has been designated Mtu SigB. A Mycobacterium leprae homologue of mysB and mtu sigB was identified in the database.
A novel repetitive DNA sequence in the sheep parasitic nematode Ostertagia circumcincta was cloned and sequenced. This 1.2-kb sequence (Oc1B) was not found in the closely related cattle parasite Ostertagia ostertagi, nor in the more distantly related sheep parasites Haemonchus contortus or Trichostrongylus colubriformis. Sequences similar to Oc1B were found at various genomic locations and contained...
A cDNA encoding a putative bovine intercellular adhesion molecule (ICAM)-3, a ligand of the leukocyte integrin LFA-1 (CD11a/CD18), was sequenced and compared with human ICAM sequences. The 1635-bp bovine sequence codes for a protein of 544 amino acids (aa). This putative bovine ICAM-3 has five immunoglobulin (Ig)-like domains similar to human ICAM-1 and ICAM-3, and belongs to the Ig gene superfamily...
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...
The mRNA differential display method was used to isolate antibacterial defense genes from Trichoplusia ni. The mRNA population in last-instar T. ni larvae injected with bacteria was compared to that of untreated larvae. Using a PCR amplified probe corresponding to an induced mRNA, we were able to clone an attacin homolog from a λ cDNA library from vaccinated larvae. The corresponding protein showed...
In a previous work, all three components of Comamonas testosteroni B-356 biphenyl (BPH)/chlorobiphenyls (PCBs) dioxygenase (dox) have been purified and characterized. They include an iron-sulphur protein (ISP BPH ) which is the terminal oxygenase composed of two subunits (encoded by bphA and bphE), a ferredoxin (FER BPH ) encoded by bphF and a reductase (RED ...
The biotin operon of Erwinia herbicola was cloned and characterized. The operon consists of five genes arranged in the order,bioABFCD . The operon is negatively regulated via the interaction of a proposed biotin repressor with an operator sequence that lies between the bioA and bioB genes. The nucleotide sequences of bioA (7,8-diaminopelargonic acid transferase),bioB (biotin synthetase) and the...
A full-length human RHAMM cDNA clone was isolated by a combination of screening a human breast cDNA expression library with the murine RHAMM 2 cDNA as well as 5 RACE and RT-PCR using messenger RNA from human breast cell line (MCF-10A). The full-length cDNA contained 725 aa that encoded an 84 kDa protein. Although the coding region of the humanRHAMM cDNA resembles the murine RHAMM v4, it has additional...
We have cloned and characterized the gene encoding inosine monophosphate dehydrogenase (IMPDH) from Pyrococcus furiosus (Pf), a hyperthermophillic archeon. Sequence analysis of the Pf gene indicated an open reading frame specifying a protein of 485 amino acids (aa) with a calculated M r of 52900. Canonical Archaea promoter elements, Box A and Box B, are located -49 and -17 nucleotides (nt),...
We have cloned and characterized the gene encoding inosine monophosphate dehydrogenase (IMPDH) from Arabidopsis thaliana (At). The transcription unit of the At gene spans approximately 1900 bp and specifies a protein of 503 amino acids with a calculated relative molecular mass (M r ) of 54 190. The gene is comprised of a minimum of four introns and five exons with all donor and acceptor splice...
The present report describes the use of computer analyses to reveal a hobo-like element in the genome of Caenorhabditis elegans. Thishobo -like sequence is 3039 bp long, contains two inverted terminal repeats of 25-27 bp and probably does not encoded a functional transposase. Sequence comparisons suggest that each transposase of hobo elements probably has a D(D/S)E motif. Thus the transposases...
A genetic analysis system of Burkholderia cepacia (Bc) was developed which included transposon mutagenesis and complementation of mutation with the cloned genes of interest. To deliver the transposon in this multidrug-resistant microorganism, two plasmids, pKN30 and pKN31, were constructed which contained Tn5 derivatives, Tn5-30Tp and Tn5-31Tp, respectively, carrying Km R and Tp R ...
Chlorocatechol 1,2-dioxygenase (CC12O) and 1,2-dichlorophenol hydroxylase (DCPH) encoding genes tfdC and tfdB are located on a 4.2-kb DNA fragment cloned from the 2,4-dichlorophenoxyacetic acid (2,4D) degrading plasmid pEST4011. The nucleotide sequences of tfdC and tfdB were determined. The DCPH is coded by a 1758-bp gene and CC12O is coded by a 762-bp gene. The deduced M r of these proteins...
Members of the CCCH zinc finger (Zf) protein family have in common two or more repeats of a novel Zf motif consisting of Cys and His residues in the form Cx 8 Cx 5 Cx 3 H [where x is a variable amino acid (aa)]. We used a degenerate polymerase chain reaction (PCR) strategy to clone members of this gene family from Saccharomyces cerevisiae. The deduced aa sequences encoded...
Two genes, 5 -orfD-orfF-3 , were found in the genome of the archaeon Methanosarcina mazei S-6 that encode the deduced proteins, OrfD and OrfF, with structural motifs typical of the nucleotide-binding components of the ABC-transporter systems of Bacteria and Eukarya. These motifs, and other similarities of OrfD and OrfF with bacterial and eukaryal counterparts, indicate that the two archaeal molecules...
Adrrenodoxin oxidoreductase (ADR) and adrenodoxin (ADX) are the two proteins involved in electron transport to mammalian mitochondrial P-450s capable of steroid modifications. The cloning and sequencing of a S. cervisiae ADR homologue (YADR) is presented here. The YADR protein sequence shares 36 and 37% of identical amino acids with human and bovine ADR respectively. The physiological role of this...
The construction of vectors for use in Schizosaccharomyces pombe using the his3 + gene as a selectable marker is described. In addition, we report the construction of a genomic library in a his3 + -containing shuttle vector to facilitate the cloning of genes by complementation of mutant function in strains defective for His3 activity.
In Escherichia coli and most other microorganisms, peptide synthesis is started at methionine start codons which are read only by N-formyl-methionine-tRNA. The formyl group is normally removed from the N-terminal Met residue of the peptide by peptide deformylase (PDF). However, it has been observed that overproduction of proteins in recombinant bacteria often yields protein products which are incompletely...
We have identified and sequenced cDNAs of the parasitic nematode Onchocerca volvulus which encode a homologue of phosphatidylethanolamine-binding proteins from mammals. These clones are also closely related to the O. volvulus Ov16 cDNA (Lobos et al., 1990). One identified cDNA clone appears to represent a partially processed precursor. These results suggest that these cDNAs are derived from a complex...
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