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We have amplified and sequenced parts of the genomes of eleven laboratory strains of bovine viral diarrhea (BVD) virus originating from North America, New Zealand and Europe. The cumulative nucleotide (nt) sequence heterogeneity of the amplified fragments located in the analysed region of the gene encoding the nonstructural protein NS3 (P80) was 24% as compared to 47% for E2 (Gp53). The nt substitutions...
The genomic sequence of Hoxa-7 (encoding the HOXa-7 homeobox protein), including the coding region (0.7 kb), flanked by a 5 -upstream region (2.8 kb), a 3 -downstream region (1 kb) and interrupted by an intron (995 bp), was determined. Northern blot analysis indicated the transcript size of Hoxa-7 to be 2.1-2.4 kb. Reverse transcription-PCR and primer extension analysis established the 5 -boundary...
We have characterized cDNA clones encoding the α 1 and β 1 subunits of Na,K-ATPase produced in the human retinal pigment epithelium (hRPE). In addition to isolating clones corresponding to known sequences of Na,K-ATPase subunits, we report hitherto unknown forms of Na,K-ATPase with unique deduced amino acid (aa) sequences in their C-termini. Truncated cDNA sequences were found for...
The 3 -terminal nucleotide (nt) sequence of the coat protein (CP)-encoding gene of the Korean isolate of cymbidium mosaic potexvirus (CyMV-K) has been determined. The sequence contained a full-length ORF coding for the viral CP. It comprises 663 nt and a polypeptide chain of 220 amino acids with a M r of 23 718. The stop codon (TGA) is followed by a 3 -untranslated region (3 -UTR) of...
We have isolated two lipoprotein lipase (LPL)-encoding cDNA (LPL) clones from a baboon cardiac cDNA library, one of which spans a region from nucleotide (nt) 705 of the coding sequence to the poly(A) tail (2.8 kb). We used reverse transcription followed by PCR (RT-PCR), and anchor-ligated rapid amplification of cDNA ends (RACE) to amplify the remaining 5 region of the LPL transcript. Sequence comparisons...
An excellent model for studying heart development in vertebrates is the cardiac non-function lethal mutant (gene c) Mexican axolotl, Ambystoma mexicanum. In order to facilitate our analyses of the mutant system, we have undertaken a search for stage-specific molecular markers during embryonic development of the axolotl. We have concentrated on homeo☐ genes 'as suitable candidates for monitoring molecular...
A novel cDNA encoding PTP (protein tyrosine phosphatase) was cloned from PC12h cells and designated as PCPTP1 (gene encoding PC12 protein Tyr phosphatase). The longest open reading frame (ORF) of this clone encodes a 656-amino-acid (aa) protein with a single PTP catalytic domain. Western blot analysis using a polyclonal Ab (antibody) raised against the cytoplasmic region of PCPTP1 detected two products,...
The amino acid (aa) sequence of rabbit opsin from rod photoreceptor cells was determined by direct aa sequencing and conceptual translation from the cDNA. The cDNA (1198 bp) containing the complete coding region encodes a 348-aa opsin protein. Of the 16 rod cell opsins that are known, rabbit opsin is most similar to human opsin (96.3% identity at the aa level).
Three differently sized mRNAs are expressed from each of two DHFR (encoding dihydrofolate reductase) alleles present in the Chinese hamster lung (CHL) cell line, DC-3F. The relative abundancy of the transcripts produced from each allele differs dramatically as a result of differential utilization of the multiple poly(A) sites present in the DHFR gene and a genetic polymorphism located within the...
We have utilized RT-PCR to clone the porcine p55TNFR cDNA, encoding the 55-kDa tumor necrosis factor receptor (TNFR), encompassing the entire coding region and most of the 3 untranslated region. PCR was performed using total cellular RNA of porcine kidney cell line 15 [PK(15)] and primers for the human p55TNFR. Since the length of the entire fragment was over 2000 bp, we fused two amplified subfragments...
Aspergillus fumigatus (Afu) and A. flavus (Afl), two causative agents of invasive aspergillosis, produce highly homologous serine proteinases. In addition, the former produces a 42-kDa metalloproteinase (MEP), whereas the latter produces a 23-kDa MEP. The cDNA and the gene encoding the 42-kDa MEP were cloned and sequenced. Here, we report the cloning of the cDNA and the gene encoding the 23-kDa...
We previously reported the identification of the human CAP and CAP2 genes which encode proteins related to the yeast adenylyl cyclase (CYR)-associated CAP protein. The rat CAP homolog, MCH1, has also been previously cloned. We have cloned a cDNA encoding the rat homolog of CAP2. Rat CAP/MCH1 and CAP2 are 63% identical to each other. Using the reverse transcription-polymerase chain reaction (RT-PCR)...
Metarhizium anisophilae (Ma) secretes a range of proteases when grown in vitro on insect cuticle. A trypsin-like serine protease, PR2, was purified from culture filtrates by anion exchange chromatography and the N-terminal sequence determined. Using oligodeoxyribonucleotide probes based on this sequence and that of the highly conserved trypsin active site, a gene was isolated from a λEMBL3 genomic...
The rotund (rn) gene in Drosophila melanogaster codes for a RacGTPase-activating protein, RnRacGAP. Cellular studies have shown that RacGAP proteins function as negative regulators of substrate Rac proteins which, in turn, control the localization and polymerization state of actin within the cell. Previous sequence analysis of rn genomic DNA and incomplete cDNA clones suggested that at least two...
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...
A full-length cDNA clone of the blotch isolate of the peanut stripe potyvirus (PStV) RNA genome was constructed downstream from the bacteriophage SP6 RNA polymerase promoter. The full-length PStV cDNA clone (PStVSF9) was sequenced and compared to the previously published sequence of PStV-B. In vitro-synthesized PStV transcripts capped with m 7 GpppG were infectious in Nicotiana benthamiana...
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...
Alu repeat sequences and other multiple copy repetitive elements are present throughout the human genome and are active in promoting recombination. It is believed that reverse transcription of transcribed Alu repeats followed by chromosomal integration has been responsible for the wide dispersion and high copy number of these sequences. During studies on the hMSH2 gene we have used RT-PCR to amplify...
A cDNA encoding a putative alcohol dehydrogenase class III (ADH) was cloned from a cDNA library constructed from 7-day larvae RNA of the marine teleost Sparus aurata. The full length cDNA is 1350 nucleotides (nt) long and contains an ORF of 1128 nt [encoding 376 amino acid (aa) residues]. Identity of 82% was found with human class III ADH (305 of 372 aa compared), and only 62% identity with a fish...
The structure and temporal expression of two Xenopus cDNAs encoding the β subunit of pyruvate dehydrogenase (XPdhE1β) have been determined. XPdhE1β was 88% homologous to mature human PdhE1β, but the putative N-terminal mitochondrial signal peptide was poorly conserved. Zygotic expression of XPdhE1β mRNA was detected at neural tube closure and increased until stage 40. RT-PCR cloning identified a...
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