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The pcp gene, encoding a pyrrolidone carboxyl peptidase (PYRase), was cloned from a λGT11 genomic library prepared from Staphylococcus aureus FDA 574 and sequenced. The pcp gene is located 740 bp downstream from cna, a gene that encodes a collagen-binding adhesin in S. aureus. S. aureus pcp encodes a 212-amino-acid (aa) polypeptide. Thepcp gene was overexpressed in Escherichia coli and the PYRase...
An 800-bp fragment of genomic DNA upstream from the origin of transcription of the mouse βB2-crystallin-encoding gene (βB2-Cry) has been isolated and its nucleotide sequence determined. Promoter fragments 275 to +30 or -110 to +30, fused to cat reporter gene, activated transcription in transiently transfected rabbit lens epithelial cells, but not in various non-lens cells. The βB2-Cry mouse promoter...
While cyclin-dependent kinases, such as CDC2 and CDK2, are key regulators of cell-cycle progression, cyclin-dependent kinase 5 (CDK5) is highly expressed in mature neurons with no evident cell-cycle regulation. The 5 -region of the mouse CDK5 gene was isolated and sequenced. The isolated clone included exons 1 through 7. The 5 -flanking region has a high G + C content. There is no TATA box around...
A COS-like monkey kidney cell line stably transfected with the plasmids pCMV gagpol-rre-r with the gag and pol genes, and pCMV rev with the rev gene of HIV-1 derived from the cDNA clone BH10, was used as a model for assessing the effectiveness of antisense (AS) constructs. A 20-mer oligodeoxyribonucleotide (oligo) phosphorothioate sequence (5 -CCG CCC CTC GCC TCT TGC CG) complementary to a portion...
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...
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...
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 have identified a second human homolog of the yeast Ste20 protein kinase family, which we designate MST2. MST2 is most similar to the previously identified MST1 protein kinase (78% identity, 88% similarity). Northern analysis indicates that MST2 mRNA is expressed at high levels in adult kidney, skeletal and placental tissues and at very low levels in adult heart, lung, liver and brain tissues...
Members of the Ras family of proteins are important components of signal transduction pathways responding to external signals and leading to changes in cell behavior. Analysis of two ras-related genes in the phylogenetically old metazoan Hydra indicates that in normal animals both genes are expressed in all body regions of the polyp. Upon head removal, however, the transcript level of one of the...
We have isolated 52 mouse cardiac troponin-T-encoding cDNA clones (TnT) by specific antibody screening of a λZAPII expression library. Sequencing data from the large sample of independent cDNAs demonstrated relationships among the expression of four alternatively-spliced exons of the cardiac TnT gene, producing seven classes of cDNAs encoding four protein isoforms differing in two variable regions...
In the bovine herpesvirus-1 (BHV-1) genome, a gene equivalent to the glycoprotein K (gK)-encoding gene of other herpesviruses was identified and sequenced. The primary translation product is predicted to comprise 338 amino acids (aa) and to exhibit a molecular mass of 37.5 kDa. It possesses characteristics typical for membrane glycoproteins including a potential cleavable signal sequence, three...
A genomic clone encoding the rice endosperm major globulin (α-globulin) with an apparent molecular mass of 26 kDa was isolated, and its nucleotide (nt) sequence and transcription start point (tsp) were determined. The tsp was identical to that of the gene encoding the wheat high-molecular-weight (HMW) glutenin subunit. The consensus -300 element and an A + T-rich sequence exist upstream from...
The reduction of nitrate to nitrite catalyzed by nitrate reductase (NR) is considered to be the rate-limiting and regulated step of nitrate assimilation, a major metabolic pathway occurring in a wide range of organisms which in turn supply the nutritional nitrogen requirements for other forms of life. Chlorella vulgaris NR mRNA levels are very responsive to changes in nitrogen source. In the presence...
We have isolated and sequenced the 5 end of the cDNA encoding the rat microtubule-associated protein 1B (MAP1B). We found that this region is highly homologous to the corresponding regions of the human [Lien et al., 22 (1994) 273-280] and mouse [Noble et al., J. Cell Biol. 109 (1989) 3367-3376] MAP1B genes. The combination of the sequence that we are presenting with the previously published sequence...
A vector is described for the expression of genomic or cDNA copies of bovine major histocompatibility complex (MHC) class I genes in transfected mouse Ltk - cells. Class I gene fragments are amplified by the polymerase chain reaction, using primers in conserved parts of exon 2 and the 3 -untranslated region of the gene. Amplified class I gene fragments can then be subcloned into the expression...
Farnesyl pyrophosphate synthetase (FPS; EC 2.5.1.10) produces the 15-carbon farnesyl pyrophosphate which is utilized in the synthesis of sterols, carotenoids, dolichols, coenzyme Q, heme a and farnesylated proteins. We have cloned this mRNA sequence from a maize endosperm cDNA library and determined the 1378-nucleotide (nt) sequence of the DNA fragment. This sequence specifies an open reading frame...
The cDNA clones encoding rabbit lung phosphatidylinositol transfer protein (PI-TP) were isolated and sequenced. The putative polypeptide consisted of 270 amino acid (aa) residues, the same as human PI-TP, but one aa residue less than the PI-TP of rat and mouse. PI-TP RNA expression in various tissues of a pregnant rabbit was analyzed by Northern blot. Brain, placenta and fallopian tube had the...
The Drosophila melanogaster try29F gene encodes a protein that shares all known features of serine proteases, like residues known to be involved in substrate specificity, catalysis and disulfide bond formation. In situ hybridization to mRNA in whole mount embryos shows that the expression of try29F is restricted to the posterior midgut during late embryogenesis.
As a first step in analyzing the function of a cdc25 homolog during the embryonic development of Patella vulgata (Pv), genomic clones encoding these stringlike proteins (Stl) were isolated and characterized. These clones belong to four groups which are derived from different regions of the Pv genome. As the sequences of Stl genes from two of these groups are almost identical, we suggest that these...
We report the complete sequence and expression of a cDNA clone (Pm3-3) encoding a cysteine protease (CysP) fromPseudotsuga menziesii [Mirb] (Pm) Franco (Douglas fir). The sequence consists of a 5 untranslated region (UTR) of 153-bp followed by an open reading frame (ORF) of 1362 bp encoding a putative mature CysP flanked by N- and C-terminal propeptides. A 364-bp 3 UTR contains multiple putative...
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