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A full-length cDNA clone coding for a 248-amino-acid tropomyosin (TM) was isolated from a Xenopus laevis (Xl) oocyte cDNA library. This TM is very similar to the members of the non-muscle (nm) TM family that includes rat TM-4, human TM30pl and chicken cardiac fibroblast FT-C. An RNase protection assay showed that the corresponding transcript is expressed ubiquitously and revealed the presence of...
Three recombinant phages containing hamster interferon-α-encoding genes (Ha Ifa) were isolated from a Ha genomic library, using a murine (Mu) Ifa probe. The phage inserts contained overlapping genomic fragments which span a total length of approx. 30 kb, on which four Ha Ifa genes are localized. The Ifa gene cluster could be assigned to hamster chromosome 2q. The nt sequences of the four Ifa genes...
We have isolated a cDNA encoding Xenopus laevis (Xl) heat-shock factor 1 (XHSF1). XHSF1, translated from the mRNA synthesized in vitro, will bind specifically to the Xl hsp70 promoter (hsp70). Microinjection of XHSF1 mRNA into Xl oocytes leads to synthesis of XHSF1 which accumulates in the nucleus and selectively activates Xl phsp70p activity at 18°C.
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...
An Aspergillus niger (An) genomic library was constructed using the promoter-trap vector, pLX2A, which contains a hygromycin B (Hy) phosphotransferase-encoding gene (hph) for selection of DNA fragments with promoter activity. This library was transformed in Escherichia coli and 80 000 colonies were obtained, 94% of which contained inserts. Transformations of plasmid DNA from the library into An resulted...
We have isolated and sequenced the mouse somatostatin receptor subtype-4-encoding gene (mSSTR4). The mSSTR4 gene encodes 384 amino acids (aa). The deduced mouse SSTR4 has 95 and 89% aa identity with the deduced rat and human SSTR4, respectively, while it has lower aa identity with the other mouse subtypes (mSSTR1, 60%; mSSTR2, 47%; mSSTR3, 42%). Using an RT-PCR technique, we show that the mSSTR4...
The chitobiase (Chb) encoding gene (chb) from Serratia marcescens (Sm) has been cloned, sequenced and expressed in Escherichia coli (Ec). Sequencing has revealed an open reading frame encoding a protein of 885 amino acids (aa). Ec cells harbouring plasmids containing chb can produce enzymatically active Sm Chb protein which is secreted into the periplasm. An efficient purification scheme using cation-exchange...
Two proteins of Aequorea victoria were molecularly engineered and produced in mammalian cells, in order to serve as specific reporters of subcellular microenvironments. Aequorin (AEQ), a Ca 2+ -sensitive photoprotein, was successfully targeted to three intracellular locations: cytosol, nucleus and mitochondria. The recombinant apoprotein, reconstituted into active AEQ by the addition...
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...
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...
The preS1 and preS2 antigens (preS Ag) of hepatitis B virus (HBV) elicit virus-neutralizing and protective antibodies which can overcome nonresponsiveness to the currently available vaccine for HBV and also carry the attachment site to HBV hepatocyte receptor. Therefore, in order to study the development of more effective vaccine and the receptor-ligand interaction, it will be helpful to obtain high-level...
The pAC92 plasmid is a direct screening cloning vector which allows positive selection of recombinant clones (re-clones). This new high-copy-number plasmid vector encodes ampicillin resistance and carries the Bacillus subtilis α-amylase (α-Amy)-encoding gene (amy) containing a multiple cloning site. The pAC92 plasmid confers to Escherichia coli transformants an amylolytic phenotype easily detected...
The aminoglycoside 3 -phosphotransferase typeVIII (APHVIII) encoding gene (aphVIII) from Streptomyces rimosus was introduced by glass-bead high-efficiency transformation into the nuclear genome of green unicellular alga Chlamydomonas reinhardtii for induction of transformants resistant to aminoglycoside antibiotics. The aphVIII structural sequence was flanked by S. rimosus regulatory sequences which...
The first evidence for a plant tRNA Lys (CUU) gene is reported. This gene is found closely linked 400 bp upstream, and on the complementary strand, of a ZmFer2 ferritin gene in the maize nuclear genome. Southern blot analysis indicates that this tRNA Lys is a member of a multigene family. This gene does not contain any intron, and exhibits classical intragenic...
The H1 histone family in mammals contains at least seven subtypes. In the past we have isolated six of the seven genes encoding these isoforms. To complete the set of the human H1 histone genes, we have designed two PCR primers deduced from a partially published sequence of the remaining histone H1 gene [Carozzi et al. (1984) Science 224, 1115-1118] and from a consensus sequence which we have derived...
We have analyzed the promoter and the coding sequences of the two homologous histone H1 o -encoding genes from Xenopus laevis, here termed H1 o -1 and H1 o -2. Both genes encode proteins of 193 amino acids and differ at just 16 amino-acid residues. Putative regulatory sequences identified in the promoter region are the same and are highly conserved. However, significant differences...
α-Sarcin is a ribonucleolytic protein secreted by the mold Aspergillus giganteus. DNA encoding α-sarcin was isolated from the host and cloned into T7 promoter based E. coli expression vectors. Using bacterial outer membrane protein A (OmpA) signal sequence, properly processed recombinant (re-) protein was secreted into the culture medium while in the absence of a signal sequence protein remained...
Lysenin, which causes contraction of rat vascular smooth muscle, is a protein that was isolated from the earthworm Eisenia foetida. A cDNA encoding lysenin was isolated by use of a partial cDNA probe that had been generated by the PCR with a primer designed by reference to an internal peptide sequence of lysenin. This clone had an ORF encoding 297 amino acid residues. The amino acid sequence deduced...
We have isolated and determined the nucleotide sequence of the gene for the fifth mouse somatostatin receptor (msst 5 ). The gene can encode a protein of 363 amino acid residues (aa). The deduced aa sequence of msst 5 has 97 and 81% identity to the rat and human sst 5 respectively, while it has lower identities with the four other mouse sst n s (msst 1 - 48%,...
A mutation strategy which utilises phage display technology and the Escherichia coli mutator strains, mutD5-FIT and XL1-RED, was applied to a Hepatitis B (HepB) specific single-chain Fv (scFv) to incorporate random mutations throughout the gene. Messenger RNA from a hybridoma producing antibodies against HepB was isolated, reverse transcribed and used as template for the production of scFv. Following...
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