Biomedical subjects

J G Shaw

Publications and source records attributed to J G Shaw.

At least 55 records · Page 3Linked to original sources

PubMed1989

Infectious in vitro transcripts from cloned cDNA of a potyvirus, tobacco vein mottling virus.

Full-length cDNA copies of tobacco vein mottling virus (TVMV) RNA were constructed downstream from bacteriophage T7 or T3 RNA polymerase promoters. The plasmids were designed to produce in vitro transcripts containing, respectively, one or two guanosine residues at the 5' terminus not derived from the TVMV sequence and a single cytidine residue at the 3' terminus following the poly(A) tail. Introduction of transcripts from either plasmid into tobacco mesophyll protoplasts resulted in the accumulation of TVMV coat protein and RNA. Neither coat protein nor viral RNA accumulated in protoplasts inoculated with linearized cDNA or with in vitro transcripts synthesized in the absence of 7-methylguanosine(5')triphospho(5')guanosine (m7GpppG). Tobacco seedlings inoculated with native TVMV RNA developed symptoms a few days before those inoculated with in vitro transcripts; however, 3 weeks after inoculation, the symptoms produced by the two inocula were indistinguishable.

Blotting, Western↗

PubMed1988

In vitro analysis of tobacco vein mottling virus NIa cistron: evidence for a virus-encoded protease.

Potential protease functions associated with the NIa nuclear inclusion protein of tobacco vein mottling virus (TVMV) were investigated. In the absence of treatments, in vitro translation of viral RNA produced various polypeptides representing each of the proposed TVMV cistrons--28K-HC-42K-CI-5.5K-NIa-NIb-CP. When viral RNA was first hybridized to DNA probes complementary to the NIa cistron, and then treated with RNase H prior to translation, a 48-kDa processing product, immunologically identified as the NIa protein, was abolished. In its place was observed a series of larger polypeptides, immunologically identified as fusion products of the cylindrical inclusion (CI) and NIa cistrons. The use of probes which permitted translation through as few as 15 nucleotide residues beyond the sequences encoding the proposed carboxyl terminus of NIa resulted in normal processing. None of the DNA probes affected an apparent cleavage between the helper component (HC) and 42K proteins. Cloned cDNA regions representing the NIa cistron and flanking sequences were inserted in transcription vectors. Translation of the in vitro transcript resulted in synthesis, not of a large fusion polyprotein, but, of a mature-sized NIa polypeptide. In vitro transcripts, lacking the 3'-most sequences that were expected to encode the protease active site of the NIa protein, were translated. These generated an apparent fusion polypeptide that reacted with antisera to both CI and NIa. The results indicate that the NIa gene product functions as a protease responsible for some but not all of the cleavage events which lead to the production of the mature forms of TVMV proteins.

Genes, Viral↗

PubMed1988

Mapping of the tobacco vein mottling virus VPg cistron.

The location of the cistron encoding the genome-linked protein (VPg) in the potyvirus tobacco vein mottling virus (TVMV) was investigated. Precipitation of 125I-labeled VPg with anti-tobacco etch virus 49K nuclear inclusion protein antiserum (which reacts with the NIa nuclear inclusion protein of TVMV) indicated that the TVMV VPg is immunologically related to NIa. Lysyl residues were found to be present at positions 2, 11, and 16 of the amino-terminal region of the VPg. A search of the TVMV polyprotein sequence for this distribution of lysyl residues revealed a unique location beginning at amino acid residue 1801, the proposed amino-terminus of the NIa protein.

Amino Acid Sequence↗

PubMed1986

The nucleotide sequence of tobacco vein mottling virus RNA.

The nucleotide sequence of the RNA of tobacco vein mottling virus, a member of the potyvirus group, was determined. The RNA was found to be 9471 residues in length, excluding a 3'-terminal poly(A) tail. The first three AUG codons from the 5'-terminus were followed by in-frame termination codons. The fourth, at position 206, was the beginning of an open reading frame of 9015 residues which could encode a polyprotein of 340 kDa. No other long open reading frames were present in the sequence or its complement. This AUG was present in the sequence AGGCCAUG, which is similar to the consensus initiation sequence shared by most eukaryotic mRNAs. The chemically-determined amino acid compositions of the helper component and coat proteins were similar to those predicted from the nucleotide sequence. Amino acid sequencing of coat protein from which an amino-terminal peptide had been removed allowed exact location of the coat protein cistron. A consensus sequence of V-(R or K)-F-Q was found on the N-terminal sides of proposed cleavage sites for proteolytic processing of the polyprotein.

Base Sequence↗

PubMed1985

Noninvasive Doppler determination of cardiac output during submaximal and peak exercise.

We compared pulsed Doppler (PD) measurements of stroke volume (SV) and cardiac output (CO) as a function of work load with previously reported values that were obtained by standard invasive methods. Suprasternal notch measurements of Doppler-shifted frequency (delta f) were obtained from the ascending aorta and SV calculated with the Doppler equation and an independent measurement of aortic diameter. Motion artifacts were minimized with the aid of a restraining table cycle ergometer. Signal aliasing was accommodated with manual summation of delta f waveforms. A total of 207 determinations were made in 10 sitting subjects exercising to exhaustion. Linear regression analysis of CO vs. work load was significant (P less than 0.001). The correlation coefficient (r = 0.95) and standard error of estimate value (1.21 1/min) were similar to values from the literature. Absolute values of CO and SV underestimated the literature values across all work loads. Technical reproducibility was assessed by comparing with paired t tests the differences between 65 duplicate serial measurements of CO and SV at rest and exercise. No significant differences (P less than 0.001) were found. We concluded that PD-determined SV and CO are reproducible and correlate linearly with work load in a manner consistent with reported invasive techniques. Thus the PD method appears suitable for use during submaximal and peak exercise.

Adult↗

PubMed1978