18 research outputs found

    Vikingar i österled

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    Information for the primers used in the qRT-PCR analysis. (DOCX 16 kb

    Molecular Cloning, Characterization and Expression Profiling of a Ryanodine Receptor Gene in Asian Corn Borer, <i>Ostrinia furnacalis</i> (Guenée)

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    <div><p>Ryanodine receptor (RyR) Ca<sup>2+</sup> release channel is the target of diamide insecticides, which show selective insecticidal activity against lepidopterous insects. To study the molecular mechanisms underlying the species-specific action of diamide insecticides, we have cloned and characterized the entire cDNA sequence of RyR from <i>Ostrinia furnacalis</i> (named as <i>Of</i>RyR). The <i>Of</i>RyR mRNA has an Open Reading Frame of 15324 bp nucleotides and encodes a 5108 amino acid polypeptide that displays 79–97% identity with other insects RyR proteins and shows the greatest identity with <i>Cnaphalocrocis medinalis</i> RyR (97%). Quantitative real-time PCR showed that the <i>Of</i>RyR was expressed at the lowest level in egg and the highest level in adult. The relative expression level of <i>Of</i>RyR in first, third and fifth-instar larva were 1.28, 1.19 and 1.99 times of that in egg. Moreover, two alternative splicing sites were identified in the <i>Of</i>RyR gene. One pair of mutually exclusive exons (a/b) were present in the central part of the predicted SPRY domain, and an optional exon (c) was located between the third and fourth RyR domains. Diagnostic PCR demonstrated that exons a and b existed in all developmental stages of <i>Of</i>RyR cDNA, but exon c was not detected in the egg cDNA. And the usage frequencies of these exons showed a significant difference between different developmental stages. These results provided the crucial basis for the functional expression of <i>Of</i>RyR and for the discovery of compound with potentially selective insect activtity.</p></div

    Primers used in cloning <i>Of</i>RyR cDNA.

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    <p>Note: R = A/G, Y = C/T, M = A/C, K = G/T, S = C/G, W = A/T, H = A/C/T, B = C/G/T, V = A/C/G, D = A/G/T, N = A/C/G/T. These primers were synthetized by BGI (Beijing, China).</p

    The usage frequencies of <i>Of</i>RyR alternative exons (a and b) in different developmental stages of O. furnacalis.

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    <p>Mutually exclusive exons were represented by yellow bars (exon a) and open bars (exon b). 1st, 3rd and 5th denotes first-instar, third-instar and fifth-instar larva. Different letters above each bar indicate statistical difference by ANOVA followed by the Duncan's Multiple Comparison test (P<0.05).</p

    The relative expression levels of <i>Of</i>RyR in different developmental stages of <i>O. furnacalis</i>.

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    <p>Data were presented as means ± SE for four independent replicates. 1st, 3rd and 5th denotes first-instar, third-instar and fifth-instar larva. Different letters above each bar indicate statistical difference by ANOVA followed by the Duncan's Multiple Comparison test (P<0.05).</p
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