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A cDNA encoding human gastric lipase (hGL) has been expressed on multicopy plasmids in the fission yeastSchizosaccharomyces pombe (Sp). Active lipase is secreted from transformants containing the hGL cDNA under the control of either the Sp adh1 promoter (Padh1) or the plant cauliflower mosaic virus (CaMV) 35S promoter. Cell-wall-associated lipase activities are greatest in the early logarithmic...
The cDNAs from a hybridoma (mAb 481.1) specific for diuron, a widely used phenylurea herbicide, were cloned into the phage display vector pComb8. Antigen-binding clones were selected by panning on diuron-hapten-BSA conjugates. The nucleotide and deduced amino-acid sequences encoding the Fab regions of the light (κ) and heavy (γ) chains were determined. The light chain was from mouse κ chain subgroup...
New intron-containing genes encoding tRNAs were discovered when genomic DNA isolated from various animal species was amplified by the polymerase chain reaction (PCR) with primers based on sequences of rabbit tRNA Lys . From sequencing analysis of the products of PCR, we found that introns are present in several genes encoding tRNA Lys in mollusks, such as Loligo...
Three cDNA fragments that encoded all but the extreme N terminus of the rat ErbB3 protein were cloned by low-stringency screening of a rat liver cDNA library with a human ERBB3 probe. The remaining 5 -end of the cDNA was generated by a reverse transcription-polymerase chain reaction method, and a single full-length rat ErbB3 cDNA was assembled. A comparison of the deduced amino acid (aa) sequences...
Toxoplasma gondii (Tg) is an obligate intracellular protozoan parasite that is an important opportunistic pathogen in humans. To provide additional tools for molecular genetic analysis, we have developed a set of vectors for DNA transformation in Tg based on selection with the antibiotic phleomycin (Ph). These new vectors rely on the flanking sequences from the parasite genes GRA1, GRA2 or SAG1...
We previously described the construction of novel hybrid proteins based on the B-subunit of cholera toxin (CTB) [Backstrom et al., Gene 149 (1994) 211-217], in which a neutralizing B-cell epitope from the third variable (V3) loop in the envelope glycoprotein gp120 from human immunodeficiency virus type 1 (HIV-1) was inserted within a surface-exposed region between amino acids (aa) 55 and 64. The...
Pancreatic cholesterol esterase (CEase) is involved in cholesterol ester hydrolysis and transport of cholesterol into intestinal cells. Isolation and sequencing of the mouse CEase cDNA indicates that the murine enzyme is very similar to the human enzyme, with the exception of the highly repeated C-terminal Pro-rich repeats. Reverse transcription-polymerase chain reaction analysis reveals that the...
A precursor, p105, for one of the subunits (p50) of the NF-κB transcription factor, plays an important role in inducible expression of diverse cellular genes. p105 also functions as a cytoplasmic inhibitor for NF-κB, and the proteolytic processing of its inhibitory C-terminal region is required for generation of active NF-κB. Here, it is reported that the human p105 C-terminal region is phosphorylated...
Race 1 of Cochliobolus carbonum (Cc) makes a cyclic tetrapeptide, HC-toxin, that is necessary for its virulence on certain genotypes of maize. The synthesis of HC-toxin is catalyzed by a 570-kDa multifunctional enzyme, HC-toxin synthetase (HTS). The gene encoding HTS (HTS1) is absent from other races of Cc and from other species ofCochliobolus . Four other unrelated filamentous fungi make cyclic...
Extrachromosomal DNA forms of Drosophila retrotransposons (RTn) and retroviruses have been extensively analyzed. However, no such analysis with plant RTn has been reported. Here, we report the analysis of extrachromosomal forms of the tobacco RTn Tto1. Tto1 is one of a few active RTn of plants and has been shown to be activated in tissue culture. Extrachromosomal circular DNA forms of Tto1, with...
A plasmid has been constructed to direct the synthesis of recombinant human growth hormone (re-hGH) in Escherichia coli as a fusion protein containing a His 6 tag at the N-terminus under the control of the T5 promoter. The re-hGH was synthesized in large amounts and accumulated in the form of inclusion bodies upon induction with IPTG. Inclusion bodies were solubilized in 6 M guanidine HCl...
A composite 3945-bp cDNA that encodes rat pyruvate carboxylase (PC) has been constructed from clones isolated from a rat liver cell cDNA library and the nucleotide sequence has been determined. The rat cDNAs open reading frame encodes a protein of 1178 amino acids that is 98.6% identical (99.0% similar) to that of mouse PC and 96.0% identical (97.8% similar) to that of human PC.
A cDNA encoding the ribosomal (r) protein L3 of the parasitic nematode Toxocara canis was isolated from a library constructed from mRNA of the infective larval stage. The encoded protein has a high degree of similarity to r-protein L3 from other organisms, including mammals, yeast, plants and bacteria. The mRNA encoding r-protein L3 is derived from a single-copy gene, and experimental evidence would...
The isolation and detailed characterization of a three-β-globin gene (GloB) haplotype in the Sprague-Dawley (S-D) rat is described. An enriched library, λSDHelib, was screened with a human GloB probe, humbg44, and from which a β minor gene, Rathbbz, was isolated, sequenced and characterized. A S-D rat GloB-specific probe, Ratbgze12, derived from the Rathbbz gene, was...
Ever since the discovery that the human tryptophanyl-tRNA synthetase (TrpRS)-encoding gene is induced by interferon (IFN) [J. Fleckner et al., Proc. Natl. Acad. Sci. USA 88 (1991) 11520-11524] and contains IFN-response regulatory elements [Frolova et al., Gene 128 (1993) 237-245], the biological rationale for this induction has remained unresolved. A survey of immune system proteins in this study...
A complementary DNA (1392 bp) encoding a protein with high homology to rat reversion-induced LIM (RIL) protein was cloned from rat hepatocytes by differential screening of a subtractive (normoxic minus hypoxic) λGEM-2 cDNA library. This cDNA clone, denoted CLP-36, encodes a 327-amino-acid (aa) protein that contained a restrictively conserved LIM (a Cys-rich domain with consensus aa sequence C-X ...
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