Molecular identification, expression and function analysis on creatine kinase in <i>Trachidermus fasciatus</i>

Liu Yingying; Chen Xuezhao; Yu Shanshan; Chai Yingmei; Lin Xiaopeng; Zhu Qian

doi:10.3969/j.issn.0253-4193.2018.12.003

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Liu Yingying, Chen Xuezhao, Yu Shanshan, Chai Yingmei, Lin Xiaopeng, Zhu Qian. Molecular identification, expression and function analysis on creatine kinase in Trachidermus fasciatus[J]. Haiyang Xuebao, 2018, 40(12): 21-30. doi: 10.3969/j.issn.0253-4193.2018.12.003

Citation:

Liu Yingying, Chen Xuezhao, Yu Shanshan, Chai Yingmei, Lin Xiaopeng, Zhu Qian. Molecular identification, expression and function analysis on creatine kinase in Trachidermus fasciatus[J]. Haiyang Xuebao, 2018, 40(12): 21-30. doi: 10.3969/j.issn.0253-4193.2018.12.003

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Molecular identification, expression and function analysis on creatine kinase in Trachidermus fasciatus

doi: 10.3969/j.issn.0253-4193.2018.12.003

School of Marine Science, Shandong University(Weihai), Weihai 264209, China

Received Date: 2018-03-13
Rev Recd Date: 2018-06-25

Abstract

Abstract

In previous study, we found that creatine kinase (CK) was one of the proteins which were up-regulated in the skin mucus of roughskin sculpin Trachidermus fasciatus post Vibrio anguillarum injection. To date, the function of this gene has not been well studied in fish. In this study, the cDNA sequence of CK gene was cloned using 3' RACE and 5'RACE techniques from roughskin sculpin, named as TfM-CK. The full-length of TfM-CK cDNA was 1 474 bp, which contained an open reading frame (ORF) of 1 146 bp encoding a polypeptide of 381 amino acids. The sequence BLASTP in NCBI indicated that TfM-CK was highly conserved. Quantitative real-time PCR (qPCR) analysis showed that TfM-CK mRNA expressed in all the detected tissues, with the highest expression in the muscle. After Vibrio anguillarum infection, TfM-CK transcripts were up-regulated significantly in the muscle, spleen, skin and head kidney, with the highest increase of 900 fold in the spleen. The kinase activity of recombinant protein of TfM-CK (rTfM-CK) was relatively high with 22.0 U/mg protein. It displayed high agglutination activity to all 4 tested bacterial including 2 Gram-negative bacteria Escherichia coli, Vibrio Anguillarum and 2 Gram-positive bacteria Staphylococcus aureus and Bacillus megaterium. These data indicate that TfM-CK participate in the fish immune response against pathogen infection. The present results may provide a theoretical basis for further investigation into the functions of fish TfM-CK and its molecular mechanisms of regulating fish immune response to pathogens.
- creatine kinase,
- Trachidermus Fasciatus,
- expression,
- innate immunity,
- bacterial agglutination

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References(23)

References

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