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Molecular cloning and expression profiling of a cold stress-related chitinase gene Mpcht19 in the desert beetle Microdera punctipennis (Coleoptera: Tenebrionidae)
Pages: 286-296
Year: Issue:  3
Journal: Acta Entomologica Sinica

Keyword:  Microdera punctipennischitinasecold stressgene expressionimmunohistochemistry;
Abstract: 【Aim】To understand the mechanisms underlying cold tolerance and to discover novel coldtolerant genes in the desert beetle Microdera punctipennis. 【Methods 】 The chitinase gene in M.punctipennis was cloned by screening cold stress transcriptome data and analyzed by bioinformatics. Its expression profiles in response to low temperature( 4℃) stress and in different adult tissues were detected by real-time fluorescence quantitative PCR and immunohistochemistry. 【Results】A chitinase gene named Mpcht19( Gen Bank accession no. : KY126382) was cloned from M. punctipennis adult.Bioinformatics analysis showed that the protein Mp CHT19 encoded by Mpcht19 belongs to the type IV insect chitinase family. It is hydrophilic,and the predicted molecular weight is 27. 18 k D with no signal peptide. Phylogenetic analysis showed that Mp CHT19 is similar to Tc CHT19 of Tribolium castaneum with the amino acid sequence identity of 34. 05%. The results of real-time fluorescent quantitative PCR showed that the expression of Mpcht19 was high in the fat body and hindgut with tissue specificity. In addition,its expression was also induced by low temperature of 4℃. Immunohistochemical analysis showed that Mp CHT19 protein was expressed in the fat body and hindgut at 4℃. 【Conclusion 】Theexpression of Mpcht19 shows the tissue specificity,and is low temperature inducible. The results may help to further investigate the relationship between chitinase function and cold tolerance of M.punctipennis.
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