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We have developed a series of vectors for easy construction of translational fusions with the pho A gene (encoding the periplasmic alkaline phosphatase, PhoA) in the three reading frames. One plasmid series carries a multiple cloning site (MCS) followed by a promoterless and leaderless 5 -truncated pho A ( pho A), which in turn is followed by a kanamycin-resistance-encoding gene (kan). Another...
The mlr-2 gene from the hyperthermophilic archaeum Pyrococcus furiosus was identified from a family of clones whose expression was influenced by the presence of maltose in the medium. The sequence of 2100 bp of DNA containingmlr -2 and its flanking regions revealed a 616-amino-acid (71 kDa) open reading frame (ORF). The ORF's initiation codon appeared 10 nt into the mlr-2 message and was not preceded...
Genomic and cDNA clones of a mcol gene encoding mini-collagen (MCOL), a nematocyst capsule protein, have been isolated from a reef-building coral, Acropora donei (Anthozoa). The gene and its flanking regions, comprising 5382 bp and covering three exons and two introns, were sequenced. Exons 2 and 3 together have an open reading frame which can encode a MCOL of 176 amino acids (aa). The coral MCOL...
A rat jun-D genomic clone was isolated from a rat liver λEMBL3 library. Sequence analysis revealed a high sequence similarity with mouse jun-D, and relatively low similarity with human and chicken jun-D. In both the trans-activation and DNA-binding domains, however, the amino-acid sequences were well conserved among rat, mouse, human and chicken.
A cDNA encoding a zebrafish Sox protein (Sry-type high-mobility-group box) was isolated and sequenced. The sequence within the HMG box is close to those of the B subfamily comprising mouse Sox-1, Sox-2, Sox-3 and Sox-14. While much of the rest of the zebrafish protein is unique, there are blocks of amino acids showing considerable identity with regions of Sox-1, -2 and -3. These domains may represent...
Two c-myc genes have been isolated from a tetraploid fish, the common carp (Cyprinus carpio). The nucleotide (nt) sequences of the two genes and their flanking regions (CAM1, 8675 bp and CAM2, 5784 bp) have been determined. Both genes contain two coding exons which are homologs of c-myc exons 2 and 3 in higher vertebrates. The nt sequence homologous to c-myc exon 1 of higher vertebrates has not...
Cell- and stage-specific transcription of the myelin basic protein (MBP)-encoding gene (Mbp) in brain is regulated by arrays of cis-acting regulatory elements positioned at the 5 -flanking region of the gene. The proximal element between nucleotides -14 to -50, termed MB1, has previously been shown to have an important role in the cell-type-specific transcription of Mbp in cells derived from the...
The 80-kDa D15 antigen (D-15-Ag) has previously been shown to be a target for protective immunity and conserved amongst typeable and nontypeable Haemophilus influenzae. Here, the gene encoding D-15-Ag is shown to encode a 797-aa polypeptide which, after cleavage of the predicted signal peptide, would have a molecular mass of 85 632 Da.
We have cloned and analyzed the chicken c-sno (cellular ski nvel) gene. The promoter region and all of the intron/exon boundaries have been sequenced. The gene is approx. 12-kb long and contains six exons, the first of which is noncoding. The amino-acid sequences encoded in this first coding exon of c-sno and c-ski are highly related; however, the remainder of these two genes appears to be unrelated...
A Caenorhabditis elegans (Ce) homologue to the eukaryotic tcp-1 gene (encoding t-complex polypeptide-1) has been mapped, isolated and sequenced. Ce tcp-1 is a single-copy gene located on chromosome II. Nucleotide sequence analysis of the gene reveals the presence of four introns in the coding region and repetitive elements upstream from the start codon. The predicted Ce TCP-1 protein displays more...
An individual strain of Neisseria gonorrhoeae may produce up to 16 different DNA methyltransferases (MTases). We have used a novel cloning system that is able to detect MTase clones in the absence of direct selection [Piekarowicz et al., Nucleic Acids Res. 19 (1991)1831-1835] to identify 14 different MTase clones. Initial characterization of these clones indicates that at least seven of these MTases...
The complete nucleotide (nt) sequences of the cDNA and gene encoding the marine mollusk Aplysia kurodai (Ak) ADP-ribosyl cyclase (ADRC) which synthesizes cyclic ADP-ribose (cADP-ribose), a second messenger for Ca 2+ mobilization from endoplasmic reticulum, were determined. Ak ADRC consists of 258 amino acids (aa) (29 kDa). It shares 86% aa sequence homology with that from A. californica,...
Here we report the nucleotide sequence of the canine interleukin-2 (IL-2)-encoding cDNA. Cloning of the canineIL-2 cDNA was achieved by the polymerase chain reaction employing, as a template, a cDNA derived from mitogen-stimulated canine splenic lymphocyte mRNA. The deduced amino acid (aa) sequence of canine IL-2 consists of 155 aa and displays 84% sequence similarity to human IL-2.
We constructed several cloning vectors, designated pGZRS-18 and pGZRS-38, which were based on an endogenousActinobacillus pleuropneumoniae (Apl) 4.3-kb plasmid. They carry the lacZα-complementation fragment and MCS from pUC18, and either the bla gene under the control of a putative Apl promoter or the Km R gene from Tn903. These vectors replicate in representative strains of Apl serotypes...
A new restriction endonuclease (ENase), SgfI, has been isolated from the bacterium Streptomyces sp. SgfI recognizes the 8-bp palindrome 5 -GCGATCGC-3 and cleaves double-stranded DNA after the T in this sequence, producing a two-base 3 overhang compatible with PvuI termini. SgfI is a rare-cutting ENase and should be useful for megabase mapping experiments.
To investigate the regulation of amphibian metallothionein(MT)-encoding genes, we have isolated and sequenced theXlMT-A gene encoding Xenopus laevis (Xl) MT-A. The gene displays an overall structure similar to that of mammalianMT , with three exons interrupted by two introns. The promoter region contains a typical TATA box and two metal regulatory elements (MRE) within the first 100 bp upstream...
The regulatory element (RE) of the human leukosialin (LS)-encoding gene, that encodes a major sialoglycoprotein of human leukocyte and platelet membranes, was used to develop a novel expression vector, pKX. The vector was constructed by cloning a RE fragment and the SV40 fragment containing polyadenylation and splicing signals betweenHin dIII and BamHI sites of the pCAT-Basic vector. The transcription...
A novel gene, SpTRK, has been isolated from DNA of the fission yeast Schizosaccharomyces pombe (Sp) by hybridization to an oligodeoxyribonucleotide (oligo) probe designed from a sequence fully conserved between the potassium transporter genes TRK1 and TRK2 of Saccharomyces cerevisiae (Sc). SpTRK is a single-copy gene located on Sp chromosome I. Nucleotide sequence analysis of the cloned gene identified...
The guinea-pig leukotriene A 4 hydrolase (LTA 4 H)-encoding cDNA was isolated from a guinea-pig lung cDNA library by cross-hybridization using a human probe. The deduced amino acid (aa) sequence consists of 611 aa (68 756 Da) and contains all twelve internal peptide and N-terminal sequences determined from the purified enzyme from guinea-pig intestine. The aa identity of the guinea-pig...
MAP30 is an anti-HIV plant protein that we have identified and purified to homogeneity from bitter melon (Momordica charantia). It is capable of acting against multiple stages of the viral life cycle, on acute infection as well as replication in chronically infected cells. In addition to antiviral action, MAP30 also possesses anti-tumor activity, topological inactivation of viral DNA, inhibition...
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