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G Langsley

Publications and source records attributed to G Langsley.

65 records · Page 4Linked to original sources
PubMed1985

Chromosome-sized DNA molecules of Plasmodium falciparum.

At least seven chromosome-sized DNA molecules (750 to 2000 kilobases in length and one fraction of undetermined molecular weight) from cultured clones and isolates of Plasmodium falciparum have been separated by pulsed-field gradient gel electrophoresis. Whereas asexual blood stages and sexual stages of the same line have identical molecular karyotypes, the length of chromosome-sized DNA molecules among different geographical isolates and several clones derived from a single patient is different. These length alterations of chromosomes are the result of DNA rearrangements that must occur unrelated to sexual differentiation.

Animals↗
PubMed1985

Characterisation of P. falciparum antigenic determinants isolated from a genomic expression library by differential antibody screening.

A genomic expression library of P.falciparum has been differentially screened with a number of immune sera. The response of 9 clones to the various sera is presented, together with the DNA sequence encoding the epitopes. All but one clone are extremely A+T rich and unlike the other P.falciparum epitopes described, are not composed of amino acid repeats. One clone, which responds specifically with a protective serum, has been analysed in detail. The epitope is carried on a 160kd antigen which is transcribed from a single gene to give a protein expressed in all of the erythrocytic forms. DNA sequence of this clone reveals it to have more than one open reading frame, only one of which is transcribed in the blood stages. The possible significance of the other open readings frames is discussed.

Amino Acid Sequence↗
PubMed1985

Gametocytogenesis and ribosomal rRNA gene organisation in the rodent malarias Plasmodium chabaudi and Plasmodium berghei.

A cloned Plasmodium berghei (ANKA) isolate was syringe passaged repeatedly to generate a line that was non-infective to Anopheles stephensi. Ribosomal gene organisation of this non-infective line was then compared to its infective ancestor. DNA was also prepared from asexual parasites and gametocytes of P. chabaudi and the arrangement of the rRNA genes of this species was studied. Although macrogametocytes have many more ribosomes than microgametocytes, this increase does not appear to stem from an amplification of the rRNA genes, as no differences either in the quantity or the arrangement of the rDNA could be detected. Furthermore, the loss of infectivity of the P. berghei gametocytes does not seem to be due to a reduction or rearrangement of sequences coding for the rRNA genes. P. chabaudi and P. berghei DNA failed to show any homology to a repetitive DNA sequence cloned from P. falciparum. We conclude that this probe, PFH8rep20, is specific for P. falciparum.

Animals↗
PubMed1984

Characterisation and translation studies of messenger RNA from the human malaria parasite Plasmodium falciparum and construction of a cDNA library.

RNA was isolated from trophozoites, schizonts and mixed populations of Plasmodium falciparum. 5% of the total was poly(A+) message, of average length 1.2 kb (10-12 kb maximum) and a poly(A) content of 10%. The mRNA fractions could be translated in vitro by reticulocyte lysates supplemented either with exogenous or P. falciparum tRNA. The patterns from two independent isolates, one cloned (T9-94) and one uncloned (K1) were virtually identical. Major translation products from 16-230 kDa have been measured. The most abundant is lactate dehydrogenase (34.8 kDa). Trophozoite mRNA codes principally for proteins of less than or equal to 93 kDa, while schizont mRNA codes for additional proteins of higher molecular mass. There are marked similarities between the in vitro translation products and proteins synthesised in vivo in synchronous cultures. A number of schizont mRNA translation products (principally those of 230, 203, 185, 170, 115, 101 and 71 kDa) are specifically precipitated without post-translational modification by sera from humans exposed to malaria. A cDNA library has been constructed in phage lambda from total poly(A+) RNA and partially characterised. About 10% of the clones derive from abundant mRNA sequences. Putative actin clones have been isolated from this library and the parasite actin mRNA sized at approx. 2.8 kb.

Actins↗
PubMed1983

Cloning and characterisation of the rRNA genes from the human malaria parasite Plasmodium falciparum.

Ribosomal DNA fragments from the human malaria parasite Plasmodium falciparum have been cloned and analysed in detail. Restriction mapping shows that the cloned fragments are different. However, they do have some similarities, in particular a small stretch of A+T-rich DNA located between the small and large subunit rRNA genes. A small rRNA gene has been mapped to this A+T-rich region. Copy number analysis reveals that each fragment is represented approximately 4 times in the genome, and implies that there are a total of 8 rRNA genes organised into at least two classes of transcription unit. Analysis of a third overlapping rDNA fragment indicates that the large subunit rRNA gene of at least one transcription unit contains an intervening sequence.

Animals↗
PubMed1983

Ribosomal DNA sequences detected in malaria parasites by cytochemical hybridization.

A procedure in which fluorochrome-labelled RNA is hybridized in situ to homologous DNA sequences was used to investigate the possible application of this method to the human malaria parasite Plasmodium falciparum. Rhodamine-labelled ribosomal RNA stained the nuclei of the parasites after cytochemical hybridization. This result demonstrates that ribosomal RNA genes can be visualised. It was estimated that the hybridization efficiency was greater than 40%.

Animals↗
PubMed1982

The establishment of genomic DNA libraries for the human malaria parasite Plasmodium falciparum and identification of individual clones by hybridisation.

The DNA of Plasmodium falciparum has been purified and fragmented with the restriction endonucleases EcoRI and HindIII. The fragments have been incorporated in vitro into derivatives of bacteriophage lambda to make libraries in which most of the parasite DNA is represented. By Southern hybridisation we have been able to recover from these libraries specific clones containing (a) repetitive DNA sequences, (b) rRNA gene(s) and (c) sequences homologous to an actin gene probe. Parasite DNA from two independent sources differs markedly in the pattern of its repetitive DNA visualised by hybridisation to our repetitive clone. By contrast, the rRNA genes of the two isolates prove to be carried on identically sized fragments.

Bacteriophage lambda↗
PubMedDate not supplied

Human antisera detect a Plasmodium falciparum genomic clone encoding a nonapeptide repeat.

Plasmodium falciparum causes malaria infections in its human host. Its wide distribution in tropical countries is a major world health problem. Before a vaccine can be produced, the identification and characterization of parasite antigens is necessary. This can be achieved by the cloning and subsequent analysis of genes coding for parasite antigens. Recently established cDNA banks allow the expression of cDNA derived from the simian parasite Plasmodium knowlesi and P. falciparum in Escherichia coli. Recombinants encoding parasite antigens have been identified by immunodetection in both banks. Two of them contain repetitive units of 11 (ref. 7) or 12 (ref. 5) amino acids. We describe here the construction of an expression bank made directly from randomly generated fragments of P. falciparum genomic DNA. We detect several clones which react strongly with human African immune sera. One clone expresses an antigenic determinant composed of occasionally degenerated repeats of a peptide nonamer.

Amino Acid Sequence↗
PubMedDate not supplied

A liver-stage-specific antigen of Plasmodium falciparum characterized by gene cloning.

The liver phase of development of malaria parasites has been studied only recently and remains poorly understood compared to the other stages such as sporozoïtes, merozoïtes and gametes. Access to liver forms of Plasmodium falciparum has been improved by the development of in vivo and in vitro propagation methods, but the yield of mature schizonts remains limited and does not allow a detailed antigenic analysis. To date, only immunofluorescence assays (IFA) have permitted a description of a species and liver-stage-specific antigen(s) (LSA; ref. 3). Monospecific antibodies to these antigens have not been obtained due either to difficulty in immunizing mice (against LSA), or to poor stability of human monoclonal antibodies. Therefore, as a means of characterizing the LSA, we used an alternative immunological approach to identify clones of the corresponding LSA genes. We describe here the isolation of a DNA sequence coding for a P. falciparum liver-stage-specific antigen composed of repeats of 17 amino-acids, which is immunogenic in man.

Adult↗