Characterisation of a repetitive DNA sequence from the malaria parasite, Plasmodium falciparum

doi:10.1016/0166-6851(86)90053-8

Molecular and Biochemical Parasitology

Volume 18, Issue 1, January 1986, Pages 89-101

https://doi.org/10.1016/0166-6851(86)90053-8 Get rights and content

Abstract

A repetitive DNA fragment cloned from the malaria parasite, Plasmodium falciparum, has been analysed. It contains a 21 base pair sequence which occurs in multiple tandem repeats. Two clusters of the same repeat are found in opposite orientations on the same DNA fragment. The repetitive DNA provides an additional way to distinguish between different strains of parasite by hybridisation to genomic blots and may serve as a species-specific probe for diagnosis.

References (43)

M. Goman et al.
The establishment of genomic DNA libraries for the human malaria parasite P. falciparum and identification of individual clones by hybridisation
Mol. Biochem. Parasitol.
(1982)
L. Franzen et al.
Analysis of clinical specimens by hybridisation with probe containing repetitive DNA from Plasmodium falciparum
Lancet
(1984)
V.K. Bhasin et al.
Variations in the organisation of repetitive DNA sequences in the genomes of Plasmodium falciparum clones
Mol. Biochem. Parasitol.
(1985)
S. Thaithong et al.
Clonal diversity in a single isolate of the malaria parasite Plasmodium falciparum
Trans. R. Soc. Trop. Med. Hyg.
(1984)
B. Fenton et al.
Protein variation in clones of Plasmodium falciparum detected by two dimensional electrophoresis
Mol. Biochem. Parasitol.
(1985)
G.E. Smith et al.
The bidirectional transfer of DNA and RNA to nitrocellulose or diazobenzyloxymethyl-paper
Anal. Biochem.
(1980)
M. Casadaban et al.
Analysis of gene control signals by DNA fusion and cloning in E. coli
J. Mol. Biol.
(1980)
J. Vieira et al.
The pUC plasmids, a M13 mp-7-derived system for insertion mutagenesis and sequencing with synthetic universal primers
Gene
(1982)
J. Messing et al.
A new pair of M13 vectors for selecting either DNA strand of double-digest restriction fragments
Gene
(1982)
L.S. Ozaki et al.
Structure of the Plasmodium knowlesi gene coding for the circumporozoite protein
Cell
(1983)

L.H.T. Van der Ploeg et al.

Structure of the growing telomeres of trypanosomes

Cell

(1984)

A.W.C.A. Cornelissen et al.

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

Mol. Biochem. Parasitol.

(1985)

D.L. Brutlag

Molecular arrangement and evolution of heterochromatic DNA

Annu. Rev. Genet.

(1980)

E.M. Southern

Base sequence and evolution of guinea-pig alpha satellite DNA

Nature

(1970)

J.G. Gall et al.

The satellite DNAs of Drosophila virilis

L. Manuelidis et al.

Homology between human and simian DNA

Nature

(1978)

N. Bone et al.

Investigation of the DNA of the human malaria parasite

R.V. Guntaka et al.

Organisation of Plasmodium falciparum genome

V. Rosario

Cloning of naturally occurring mixed infections of malaria parasites

Science

(1981)

W. Trager et al.

Human malaria parasites in continuous culture

Science

(1976)

P.W.J. Rigby et al.

Labelling deoxribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase

J. Mol. Biol.

(1977)

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^∗: Present address: Department of Immunology, Institut Pasteur, 25/28 Rue de Docteur Roux, 75015 Paris Cedex 15 France.

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Characterisation of a repetitive DNA sequence from the malaria parasite, Plasmodium falciparum

Abstract

Mol. Biochem. Parasitol.

Lancet

Mol. Biochem. Parasitol.

Trans. R. Soc. Trop. Med. Hyg.

Mol. Biochem. Parasitol.

Anal. Biochem.

J. Mol. Biol.

Gene

Gene

Cell

Cell

Mol. Biochem. Parasitol.

Molecular arrangement and evolution of heterochromatic DNA

Annu. Rev. Genet.

Base sequence and evolution of guinea-pig alpha satellite DNA

Nature

The satellite DNAs of Drosophila virilis

Homology between human and simian DNA

Nature

Investigation of the DNA of the human malaria parasite

Organisation of Plasmodium falciparum genome

Cloning of naturally occurring mixed infections of malaria parasites

Science

Human malaria parasites in continuous culture

Science

Labelling deoxribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase

J. Mol. Biol.