Cloning and expression pattern of heat shock protein 90 gene from planarian Dugesia japonica

doi:10.3969/j.issn.0529-1356.2013.05.009

Abstract

Abstract:

Objective To clone heat shock protein 90 gene from planarianDugesia japonica (Djhsp90), and to examine its expression pattern in response to different stressors. Methods The full-length cDNA and genomic DNA of Djhsp90 was cloned using PCR methods, and its sequence was analyzed by the bioinformatics software, and its expression pattern in response to different stressors was detected by the fluorescent Real-time RT PCR. Results The full-length cDNA of Djhsp90 was 2354 bp containing an open reading frame (ORF) of 2148 bp, which encoded a polypeptide of 715 amino acids with five signatures motifs and the C-terminal consensus MEEVD of eukaryotic HSP90 family. The ORF sequence of genomic DNA was sequenced, and found that only one intron (48 bp) existed in Djhsp90 gene structure. The 5’ and 3’ putative splicing site followed the typical “GT-AG” rule. Djhsp90 expression began to rise at 1 day following the amputation of planarians, reached a peak at day 2 and decreased gradually at day 3 to the normal level. During the prolonged starvation, Djhsp90 expression remained a relative higher level in contrast to the controls. We verified that elevating the culture temperature strongly induced Djhsp90 expression, and the most sensitive induction occurred in the head fragment of planarians. Conclusion We have successfully cloned hsp90 gene from planarianDugesia japonica, and verified that amputation, starvation and thermal stress can induce the up-regulation of Djhsp90.

Key words: Heat shock protein 90, Gene cloning, Real-time PCR, Planarian

CLC Number:

Q812

MA Ke-xue FENG Cheng-cheng DOU He CHENG Fang-fang CHEN Guang-wen* LIU De-zeng. Cloning and expression pattern of heat shock protein 90 gene from planarian Dugesia japonica[J]. AAS, 2013, 44(6 ): 778-783.

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