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Cloning and expression profiling in response to low temperature of Ras GTPase-activating protein gene MpRas GAP in the desert beetle Microdera punctipennis( Coleoptera: Tenebrionidae)
Pages: 367-374
Year: Issue:  4
Journal: Acta Entomologica Sinica

Keyword:  Microdera punctipennislow temperature stressRas GTPase-activating protein(Ras GAP)gene cloningmRNA levelreal-time quantitative PCR;
Abstract: 【Aim 】Mitogen activated protein kinase( MAPK) cascade is one of the important signal transduction pathways in cells. Ras GTPase-activating protein( Ras GAP) gene Ras GAP and c-Jun Nterminal kinase( JNK) gene JNK are an up-stream gene and a down-stream gene in MAPK pathway,respectively. This study aims to determine the expression profiles of Ras GAP and JNK in the desert beetle Microdera punctipennis in response to low temperature. 【Methods】A full length c DNA of Ras GAP from M. punctipennis was cloned. The deduced amino acid sequence was analyzed,and the phylogynetic tree was constructed. The expression profiles of Ras GAP and JNK in M. punctipennis exposed to low temperatures were detected by using real-time quantitative PCR. 【Results】The ORF of Ras GAP c DNA from M. punctipennis is 2 523 bp in length,and was named as MpRas GAP( Gen Bank accession no. :KM677930),encoding a polypeptide of 840 amino acids with the molecular weight of 96. 594 k Da. The encoded protein MpRas GAP belongs to the Ras GAP super family. Homology analysis showed that MpRas GAP shares 89% amino acid sequence identity with Ras GAP from Tribolium castaneum. When M.punctipennis adults were exposed to 4℃ and- 4℃ for 1 h,the mRNA levels of MpRas GAP were significantly upregulated as compared with that at room temperature( 25℃). When M. punctipennis adults were exposed to 4℃ for 3 h or- 4℃ for 1 h,the mRNA levels of Mp JNK were also elevated significantly. 【Conclusion】The results of this study demonstrate that the expression of both MpRas GAPand Mp JNK in M. punctipennis can be induced by low temperature. Our results will help to further study the role of MAPK pathway in the desert insect under low temperature.
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