CLONING AND EXPRESSION ANALYSIS OF HEAT SHOCK PROTEIN 60 GENE FROM HYRIOPSIS CUMINGII

WU Sheng-Nan; LIU Da-Wei; LIU Yi; HU Bao-Qing; ZHANG Jian-Qiang; WANG Yan; WANG Ting

doi:10.7541/2014.134

Volume 38 Issue 5

Sep. 2014

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ACTA HYDROBIOLOGICA SINICA > 2014 > 38(5): 897-902. > DOI: 10.7541/2014.134

WU Sheng-Nan, LIU Da-Wei, LIU Yi, HU Bao-Qing, ZHANG Jian-Qiang, WANG Yan, WANG Ting. CLONING AND EXPRESSION ANALYSIS OF HEAT SHOCK PROTEIN 60 GENE FROM HYRIOPSIS CUMINGII[J]. ACTA HYDROBIOLOGICA SINICA, 2014, 38(5): 897-902. DOI: 10.7541/2014.134

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CLONING AND EXPRESSION ANALYSIS OF HEAT SHOCK PROTEIN 60 GENE FROM HYRIOPSIS CUMINGII

College of Life Sciences, Jiangxi Normal University, Nanchang 330022, China;
2. School of Life Sciences and Food Engineering, Nanchang University, Nanchang 330031, China

Received Date: August 10, 2013
Rev Recd Date: March 04, 2014
Published Date: September 24, 2014

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Abstract

Abstract

Heat shock protein 60 (HSP60) is an important molecular chaperone protein that mainly exists in the mito-chondria of organism. In the present study, the cDNA sequence of Hyriopsis cumingii HSP60 (hcHSP60) was cloned by 3' rapid amplification of cDNA ends methods (3'-RACE) based on a long chain sequence of hcHSP60 (2629 bp) and was determined by high flux sequencing for transcriptome of H. cumingii blood cells, and its expression in the different tissues was detected by using reverse transcription-polymerase chain reaction (RT-PCR). Results showed that the full-length cDNA of hcHSP60 was 2807 bp that contains an open reading frame of 1707 bp, encoding a protein of 568 amino acid residues with 61.04 ku of predicted molecular weight and 5.63 of the theoretical isoelectric point, which was predicted to have no signal peptide and transmembrane helices. The deduced amino acid sequence of hcHSP60 shares the highest identity (82%) with HSP60 of Biomphalaria glabrata, and the phylogenetic analysis demonstrated that they were clustered in a same clade. The results of RT-PCR indicated that hcHSP60 was constitutively expressed in all 5 examined tissues of H. cumingii with the highest expression in hepatopancreas. The expression of hcHSP60 in all de-tected tissues was up-regulated obviously by higher water temperature, suggesting that it may play an important role in the stress response against heat.
- Hyriopsis cumingii,
- Heat shock protein 60,
- Sequence analysis,
- Expression

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CLONING AND EXPRESSION ANALYSIS OF HEAT SHOCK PROTEIN 60 GENE FROM HYRIOPSIS CUMINGII

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