1984
DOI: 10.1126/science.6330899
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Mung Bean Nuclease Cleaves Plasmodium Genomic DNA at Sites Before and After Genes

Abstract: Mung bean nuclease was found to cut the genomic DNA of the malaria parasite Plasmodium at positions before and after genes but not within gene-coding regions. This cleavage, which had nearly the preciseness of a restriction nuclease, required controlled conditions in the presence of formamide. Southern blot analysis showed that the coding areas for Plasmodium actin, circumsporozoite protein, histidine-rich protein, ribosomal RNA's, and tubulin are each cleaved from genomic DNA to yield a single major band on a… Show more

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Cited by 127 publications
(65 citation statements)
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“…To provide as complete a representation of genes as possible, and to minimize bias towards speci®c sequences, a mung bean nuclease genomic library was used. Mung bean nuclease preferentially cuts malarial DNA in regions¯anking coding regions (McCutchan et al, 1984;Vernick and McCutchan, 1998). Such digestion was expected to capture long stretches of unique coding regions and avoid over-representation of¯anking sequences or introns on the array.…”
Section: Array Constructionmentioning
confidence: 99%
“…To provide as complete a representation of genes as possible, and to minimize bias towards speci®c sequences, a mung bean nuclease genomic library was used. Mung bean nuclease preferentially cuts malarial DNA in regions¯anking coding regions (McCutchan et al, 1984;Vernick and McCutchan, 1998). Such digestion was expected to capture long stretches of unique coding regions and avoid over-representation of¯anking sequences or introns on the array.…”
Section: Array Constructionmentioning
confidence: 99%
“…Recombinant phage were prepared with mung bean nuclease-sheared DNA (13,18,19) ligated into alkaline phosphatase-treated Xgtll (20). Phage producing malaria antigens were identified with a pool of sera from Aotus monkey A076.…”
mentioning
confidence: 99%
“…It has not proved as big a problem with the extraction and characterization of T. gondii RNA (Johnson, McDonald and Illana, 1986) and cDNA libraries have been produced and recombinant plasmids encoding T. gondii antigens have been identified (Johnson, Illana and McDonald, unpublished data). However, the development of new techniques such as mung bean nuclease excision of whole intact genes (McCutchan et al, 1984b) makes this method a very desirable alternative to cDNA production for recombinant vaccine development. To produce a T. gondii genomic library requires reasonable amounts of purified parasite DNA, and we bave used ultracentrifugation in a CsCl/Hoechst 33258 gradient to obtain this.…”
Section: Discussionmentioning
confidence: 99%