Linked Life Data

10219298

Source:http://linkedlifedata.com/resource/pubmed/id/10219298

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pubmed-article:10219298pubmed:abstractTextDifferential mRNA display was used to detect differences in gene expression between mock-infected and Cryptosporidium parvum-infected human adenocarcinoma cells. A reproducible band present only in C. parvum-infected cells, ddHC-10 was isolated and cloned. Northern blot analysis was used to confirm the differential expression of the HC-10 mRNA. As differential mRNA display does not differentiate between parasite and host mRNAs, Southern blot analysis was used to demonstrate that ddHC-10 represented a C. parvum gene. Northern blot analysis demonstrated that HC-10 mRNA is expressed by sporozoites prior to invasion of host cells. Screening of a C. parvum genomic library identified 2 different genomic clones, HC-10-13C and HC-10-6C. The combined genomic sequence contained a predicted open reading frame of 2,952 base pairs (bp), coding for a protein of 984 amino acids with a predicted molecular weight of approximately 106 kDa. Reverse transcription polymerase chain reaction mapping of the HC-10 transcript demonstrated that the HC-10 gene lacks introns, and the approximately 4,789-bp mRNA contains relatively large 5' (approximately 1,390-bp) and 3' (approximately 440-bp) untranslated regions. The predicted polypeptide contained a high proportion of polar amino acids, with the most abundant amino acids being serine (10.5%), threonine (9.8%), and cysteine (7.6%). The C-terminal region of the predicted polypeptide is characterized by a threonine-rich region containing multiple repeats of the sequence TTTTRP. This repeat motif is similar to that found in the mucin-like genes of vertebrates and lower eukaryotes that have been shown to play important roles in cell-cell interactions in multicellular organisms and invasion of host cells by unicellular parasites.lld:pubmed
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pubmed-article:10219298pubmed:dateRevised2007-11-15lld:pubmed
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pubmed-article:10219298pubmed:articleTitleDifferential mRNA display cloning and characterization of a Cryptosporidium parvum gene expressed during intracellular development.lld:pubmed
pubmed-article:10219298pubmed:affiliationUniversity of Minnesota, St. Paul 55108, USA.lld:pubmed
pubmed-article:10219298pubmed:publicationTypeJournal Articlelld:pubmed
pubmed-article:10219298pubmed:publicationTypeResearch Support, U.S. Gov't, P.H.S.lld:pubmed
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