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J A Napier

Publications and source records attributed to J A Napier.

At least 55 records · Page 3Linked to original sources
PubMed1995

Isolation of a cDNA encoding a cytochrome b5 specifically expressed in developing tobacco seeds.

Polymerase chain reaction (PCR) was used to amplify transcripts encoding cytochrome b5 from cDNA synthesised from RNA isolated from developing seeds of tobacco (Nicotiana tabacum L.). The sequence of the amplified products indicated that the clones encoded a second form of tobacco cytochrome b5, different from that previously characterised (Smith et al. 1994, Plant Mol Biol 25:527-537). Rapid amplification of cDNA ends (RACE)-PCR was used to amplify the 5' and 3' ends of the transcript. Northern blotting and RNAse protection assays of RNA samples isolated from different tobacco tissues indicated that this second cytochrome b5 form was expressed only in developing seeds. Therefore, it seems likely that this message is the product of a tobacco cytochrome b5 gene specifically expressed in seeds.

Amino Acid Sequence↗
PubMed1994

Expression of a biologically active plant cytochrome b5 in Escherichia coli.

Cytochrome b5 from tobacco (Nicotiana tabacum) was expressed in Escherichia coli using a T7 polymerase/promoter system as described by Studier, Rosenberg, Dunn and Dubendorff (1990) (Methods Enzymol. 185, 60-89). Transformed cells were red in colour and accumulated cytochrome b5 to a level of around 30% of the total cell protein. The purified cytochrome had oxidized, reduced and low-temperature absorbance spectra characteristic of plant microsomal cytochrome b5, and exhibited a c.d. spectrum resembling that of a mammalian cytochrome b5. The recombinant protein appeared to be correctly assembled and biologically active, being reduced by NADH in the presence of microsomal membranes prepared from the developing seeds of sunflower (Helianthus annuus). Inhibition of haem synthesis in the transformed E. coli cells expressing cytochrome b5, by the use of gabaculin or succinylacetone, prevented the assembly of the cytochrome b5 holoprotein but had little effect on the accumulation of cytochrome apoprotein. The recombinant protein expressed in E. coli therefore has the biochemical features of the higher-plant cytochrome b5 and can be used in studies of plant microsomal oxidation/reduction reactions.

Animals↗
PubMed1994

Tobacco cytochrome b5: cDNA isolation, expression analysis and in vitro protein targeting.

A full-length clone encoding cytochrome b5 has been isolated from a tobacco leaf cDNA library in lambda gt11 by PCR using degenerate primers. This cDNA encodes a protein of 139 residues which exhibits a high degree of homology to other cytochrome b5s, the message for which is expressed predominantly in developing seeds and in pigmented flower tissue. In the developing tobacco seed the mRNA is abundant at very early stages (< 10 days after flowering). Southern analysis indicated that more than one gene encodes cytochrome b5 in the tobacco genome. In vitro transcription and translation studies of the cDNA indicated that the protein inserts into the ER membrane by a non-SRP-mediated pathway and that the C-terminus of the protein is required for targeting and insertion.

Amino Acid Sequence↗
PubMed1992

Import and processing of the precursor of the Rieske FeS protein of tobacco chloroplasts.

cDNA clones encoding the precursor of the Rieske FeS protein of tobacco chloroplasts have been characterised and shown to derive from two different genes. The 5' ends of the corresponding transcripts have been cloned using primer extension and PCR. The nucleotide sequences of the cDNAs (and their 5' extensions) predict precursors for the tobacco proteins which differ in 4 amino acid residues out of a total of 228 residues and show high homology with the pea and spinach precursors. The tobacco precursor proteins contain N-terminal presequences of 49 amino acid residues which lack 17 amino acid residues present at the N-terminus of the spinach presequence. The 26 kDa precursor obtained by transcription and translation of one of these cDNAs in vitro was efficiently imported and correctly processed to the mature 20 kDa protein by isolated pea or tobacco chloroplasts. The precursor was also processed to its mature size by a peptidase present in the stroma of chloroplasts.

Amino Acid Sequence↗
PubMed1992

Import and processing of the precursor form of the gamma subunit of the chloroplast ATP synthase from tobacco.

A cDNA clone encoding the complete precursor of the gamma subunit of chloroplast ATP synthase has been isolated from a tobacco (Nicotiana tabacum) leaf cDNA library in lambda gt11. The 1.4 kb insert encodes a polypeptide of 377 amino acid residues, of which 55 residues constitute an N-terminal presequence and 322 residues make up the mature gamma subunit. Hybridisation of the cDNA to Southern blots of tobacco genomic DNA indicates the presence of two genes in the haploid genome. Transcription and translation of the cDNA in vitro produced a protein of 41 kDa which was imported by isolated pea chloroplasts and processed to the mature 36 kDa subunit. The gamma subunit precursor was processed to the mature size by a processing peptidase of 180 kDa present in pea stromal extracts.

Amino Acid Sequence↗
PubMed1992

Import and processing of the precursor of the delta subunit of tobacco chloroplast ATP synthase.

A cDNA clone encoding the complete precursor of the delta subunit of chloroplast ATP synthase has been isolated from a tobacco (Nicotiana tabacum) leaf cDNA library in lambda gt11. The 880 bp insert encodes a polypeptide of 248 amino acid residues, of which 61 residues constitute an N-terminal presequence and 187 residues make up the mature delta subunit. Transcription and translation of the cDNA in vitro produced a protein of 29 kDa which was imported by isolated pea chloroplasts and processed to the mature 20 kDa subunit. The delta subunit precursor was processed to the mature size by a processing peptidase present in pea stromal extracts. Hybridisation of the cDNA to Southern blots of tobacco genomic DNA suggests the presence of two genes in the haploid genome.

Amino Acid Sequence↗
PubMed1992

Import of the precursor of the chloroplast Rieske iron-sulphur protein by pea chloroplasts.

cDNA clones encoding the precursor of the Rieske FeS protein of the chloroplast thylakoid membrane have been isolated from a pea leaf cDNA library in lambda gt 11, following screening with antibodies to purified pea chloroplast Rieske FeS protein. The longest cDNA insert of 880 bp encodes a polypeptide of 230 amino acid residues, of which 50 residues constitute an N-terminal cleavable presequence and 180 residues make up the mature protein. Transcription and translation of the cDNA in vitro produced a polypeptide of 26 kDa which was efficiently imported by isolated pea chloroplasts and processed to the mature 20 kDa protein. Southern hybridisation to pea genomic DNA indicated the presence of a single gene encoding the Rieske FeS protein in the haploid genome.

Amino Acid Sequence↗
PubMed1992

Chloroplast import of the precursor of the gamma subunit of pea chloroplast ATP synthase.

A cDNA clone encoding the complete precursor of the gamma subunit of the pea chloroplast ATP synthase has been isolated from a pea leaf cDNA library in lambda gt 11 following detection with antibodies to the purified gamma subunit. The cDNA insert of 1.4 kbp is smaller than transcripts of about 1.6 kb detected by northern hybridisation of RNA from both light- and darkgrown pea leaves. The cDNA encodes a polypeptide of 376 amino acid residues, of which 52 residues constitute an N-terminal presequence and 324 residues make up the mature protein. Transcription and translation of the cDNA in vitro produced a protein of 42 kDa, which was imported by isolated pea chloroplasts and processed to the mature 36 kDa subunit.

Adenosine Triphosphatases↗
PubMed1992

Guidelines for the use of fresh frozen plasma. British Committee for Standards in Haematology, Working Party of the Blood Transfusion Task Force.

Fresh frozen plasma should only be used to treat bleeding episodes or prepare patients for surgery in certain defined situations. Definite indications for the use of FFP: 1. Replacement of single coagulation factor deficiencies, where a specific or combined factor concentrate is unavailable. 2. Immediate reversal or warfarin effect. 3. Acute disseminated intravascular coagulation (DIC). 4. Thrombotic thrombocytopenic purpura (TTP). Conditional uses: FFP only indicated in the presence of bleeding and disturbed coagulation: 1. Massive transfusion. 2. Liver disease. 3. cardiopulmonary bypass surgery. 4. Special paediatric indications. No justification for the use of FFP: 1. Hypovolaemia. 2. Plasma exchange procedures. 3. 'Formula' replacement. 4. Nutritional support. 5. Treatment of immunodeficiency states.

Adult↗
PubMed1991

The crossmatch.

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Blood Grouping and Crossmatching↗