Cloning and comparison of repeated DNA sequences from the human filarial parasite Brugia malayi and the animal parasite Brugia pahangi.
Identifieur interne : 009517 ( Ncbi/Checkpoint ); précédent : 009516; suivant : 009518Cloning and comparison of repeated DNA sequences from the human filarial parasite Brugia malayi and the animal parasite Brugia pahangi.
Auteurs : L A Mcreynolds ; S M Desimone ; S A WilliamsSource :
- Proceedings of the National Academy of Sciences of the United States of America [ 0027-8424 ] ; 1986.
Descripteurs français
- KwdFr :
- ADN (génétique), Animaux, Animaux sauvages, Brugia (génétique), Chats, Chiens, Clonage moléculaire, DNA restriction enzymes, Filariose lymphatique (médecine vétérinaire), Filariose lymphatique (parasitologie), Haplorhini, Humains, Hybridation d'acides nucléiques, Séquence nucléotidique, Séquences répétées d'acides nucléiques.
- MESH :
- génétique : ADN, Brugia.
- médecine vétérinaire : Filariose lymphatique.
- parasitologie : Filariose lymphatique.
- Animaux, Animaux sauvages, Chats, Chiens, Clonage moléculaire, DNA restriction enzymes, Haplorhini, Humains, Hybridation d'acides nucléiques, Séquence nucléotidique, Séquences répétées d'acides nucléiques.
English descriptors
- KwdEn :
- MESH :
- chemical , genetics : DNA.
- genetics : Brugia.
- parasitology : Elephantiasis, Filarial.
- veterinary : Elephantiasis, Filarial.
- Animals, Animals, Wild, Base Sequence, Cats, Cloning, Molecular, DNA Restriction Enzymes, Dogs, Haplorhini, Humans, Nucleic Acid Hybridization, Repetitive Sequences, Nucleic Acid.
Abstract
A 320-base-pair repeated sequence was observed when DNA samples from the filarial parasites Brugia malayi and Brugia pahangi were digested with the restriction endonuclease Hha I. A 640-base-pair dimer of the repeated sequence from B. malayi was inserted into the plasmid pBR322. When dot hybridization was used, the copy number of the repeat in B. malayi was found to be about 30,000. The 320-base-pair Hha I repeated sequences are arranged in direct tandem arrays and comprise about 12% of the genome. B. pahangi has a related repeated sequence that cross-hybridizes with the cloned B. malayi Hha I repeat. Dot hybridization with the cloned repeat shows that the sequence is present in B. malayi and in B. pahangi but not in four other species of filarial parasites. The cloned repeated DNA sequence is an extremely sensitive probe for detection of Brugia in blood samples. Hybridization with the cloned repeat permits the detection of DNA isolated from a single parasite in an aliquot of blood from animals infected with B. malayi. There are differences in the restriction sites present in the repeated sequences that can be used to differentiate between the two Brugia species. The B. malayi repeated DNA sequence is cleaved by Alu I and Rsa I but the B. pahangi sequence is not. A comparison of repeated sequences between the two species by DNA sequence analysis indicates that some regions of individual repeats are over 95% homologous, while other short regions are only 60-65% homologous. These differences in DNA sequence will allow the construction of species-specific hybridization probes.
Url:
PubMed: 3003750
PubMed Central: 322952
Affiliations:
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PMC:322952Le document en format XML
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<term>Nucleic Acid Hybridization</term>
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<term>Chiens</term>
<term>Clonage moléculaire</term>
<term>DNA restriction enzymes</term>
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<term>Séquence nucléotidique</term>
<term>Séquences répétées d'acides nucléiques</term>
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<front><div type="abstract" xml:lang="en"><p>A 320-base-pair repeated sequence was observed when DNA samples from the filarial parasites Brugia malayi and Brugia pahangi were digested with the restriction endonuclease Hha I. A 640-base-pair dimer of the repeated sequence from B. malayi was inserted into the plasmid pBR322. When dot hybridization was used, the copy number of the repeat in B. malayi was found to be about 30,000. The 320-base-pair Hha I repeated sequences are arranged in direct tandem arrays and comprise about 12% of the genome. B. pahangi has a related repeated sequence that cross-hybridizes with the cloned B. malayi Hha I repeat. Dot hybridization with the cloned repeat shows that the sequence is present in B. malayi and in B. pahangi but not in four other species of filarial parasites. The cloned repeated DNA sequence is an extremely sensitive probe for detection of Brugia in blood samples. Hybridization with the cloned repeat permits the detection of DNA isolated from a single parasite in an aliquot of blood from animals infected with B. malayi. There are differences in the restriction sites present in the repeated sequences that can be used to differentiate between the two Brugia species. The B. malayi repeated DNA sequence is cleaved by Alu I and Rsa I but the B. pahangi sequence is not. A comparison of repeated sequences between the two species by DNA sequence analysis indicates that some regions of individual repeats are over 95% homologous, while other short regions are only 60-65% homologous. These differences in DNA sequence will allow the construction of species-specific hybridization probes.</p>
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