1989
DOI: 10.1016/0166-6851(89)90008-x
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Rapid changes in the expression of a gene encoding a calcium-binding protein in Schistosoma mansoni

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Cited by 53 publications
(40 citation statements)
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“…Isolation of SmRK-1 cDNA and Sequence Analysis-Total RNA was isolated from adult S. mansoni parasites using RNAzol B (Tel-Test, Inc.) and first-strand cDNA synthesis performed using M-MLV reverse transcriptase (Life Technologies, Inc.) and an oligo(dT) [12][13][14][15][16][17][18] primer. A pair of degenerate primers (forward primer: 5Ј-AAACCAGCTATTGCT-CAYMGNGA-3Ј; reverse primer: 5Ј-TCTGGAGCCATATAACGTTTT-GTNCCNAC-3Ј; corresponding to conserved sequences from subdomains VIB and VIII of other RSTK kinase domains, respectively) was used to amplify schistosome RSTK sequences from parasite cDNA by PCR.…”
Section: Methodsmentioning
confidence: 99%
See 1 more Smart Citation
“…Isolation of SmRK-1 cDNA and Sequence Analysis-Total RNA was isolated from adult S. mansoni parasites using RNAzol B (Tel-Test, Inc.) and first-strand cDNA synthesis performed using M-MLV reverse transcriptase (Life Technologies, Inc.) and an oligo(dT) [12][13][14][15][16][17][18] primer. A pair of degenerate primers (forward primer: 5Ј-AAACCAGCTATTGCT-CAYMGNGA-3Ј; reverse primer: 5Ј-TCTGGAGCCATATAACGTTTT-GTNCCNAC-3Ј; corresponding to conserved sequences from subdomains VIB and VIII of other RSTK kinase domains, respectively) was used to amplify schistosome RSTK sequences from parasite cDNA by PCR.…”
Section: Methodsmentioning
confidence: 99%
“…To detect SmRK-1 transcripts, the forward primer 5Ј-TCTGAAGTCTA-GAAACATCTTAG-3Ј and reverse primer 5Ј-GATCTGAGTTTGAATTA-ACTTC-3Ј (corresponding to nucleotides 1314 to 1336 and 1414 to 1435 of the SmRK-1 cDNA sequence, respectively) were used to amplify a 121-bp segment of the SmRK-1 cDNA sequence. As a control, a 230-bp segment of the message for a S. mansoni calcium-binding protein (CaBP) which is preferentially expressed in larval stages (14) was amplified using the forward primer 5Ј-TTTCACTGTATTGCATTA-GAATGG-3Ј and reverse primer 5Ј-TTATGAACATAAAACATCAAG-GAG-3Ј (corresponding to nucleotides Ϫ20 to ϩ4 and ϩ278 to ϩ301 of the CaBP genomic sequence).…”
mentioning
confidence: 99%
“…The viability of isolated tails was further supported by analyses for the mRNA of the cercarial-specific calcium-binding protein (CaBP). It was previously shown in our laboratory that CaBP mRNA was undetectable in sporocysts, it was abundant in cercariae and 3 h schistosomula, detected in very low amounts in 24 h schistosomula, and not detected in adult worms (Ram et al, 1989). Northern blots, hybridized with the CaBP cDNA, showed large amounts of the CaBP mRNA in isolated tails as in intact cercariae and isolated bodies kept for 3 h at 23 "C and 37 "C (data not shown; see Neumann, 1992).…”
Section: Increased Expression Of the Hsp70 Gene During Cercariae/schimentioning
confidence: 99%
“…A cDNA library of S. mansoni adult worm was constructed in the Agtll expression vector, as described (Maniatis et al, 1982;Ram et al, 1989). Clone-3M (0.7 kb) was isolated by screening the library with sera of mice infected with S. mansoni.…”
Section: Dna Probe Antigens and Antiseramentioning
confidence: 99%
“…The other calcium-binding protein is found almost exclusively in the cercariae stage. This gene seems to be turned on within a short period of time after the release of the cercariae into the water and shut off when the cercariae transforms into schistosomula (Ram et al 1989). The calcium-binding proteins are interesting because several metabolic and physiological events are triggered by calcium ions with the help of these molecules (Goodman et al 1979).…”
Section: Discussionmentioning
confidence: 99%