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Molecular cloning and analysis of function of nucleoside diphosphate kinase (NDPK) from the scallop Chlamys farreri

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Abstract

Nucleoside diphosphate kinase (NDPK) is a key metabolic enzyme that catalyzes the synthesis of non-adenine nucleoside triphosphate (NTP) by transferring the terminal phosphate between nucleoside diphosphate (NDP) and NTP. NDPK regulates a variety of eukaryotic cellular activities including cell proliferation, development, and differentiation. The ndpk cDNA was cloned from the hemocytes of the scallop Chlamys farreri and designated Cf-ndpk. The full-length sequence of Cf-ndpk consists of 715 bp encoding a polypeptide of 153 amino acids with a calculated molecular mass of 16927.52 daltons and pI of 7.64. The mRNA expression and distribution of Cf-ndpk in both bacterially challenged and unchallenged scallops were studied by Northern blotting and in situ hybridization. The results showed that Cf-ndpk transcripts were present in hemocytes, gill, adductor muscle, mantle, digestive gland, foot, and gonad, and the expression level increased in hemocytes after bacterial challenge. Recombinant Cf-NDPK expressed in Escherichia coli could transfer the terminal phosphate between UDP and ATP. The Cf-NDPK protein was present in all tested tissues including foot, adductor muscle, digestive gland, gonad, mantle, gill, and hemolymph. It was up-regulated in hemolymph after bacterial challenge. Taken together, these results suggest that NDPK may play roles in the innate immune response of scallop.

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Abbreviations

EST:

expression sequence tags

IPTG:

isopropyl-1-thio-β-D-galactopyranoside

NDPK:

nucleoside diphosphate kinase

Cf-NDPK:

nucleoside diphosphate kinase from the scallop Chlamys farreri

RT-PCR:

reverse transcription polymerase chain reaction

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Authors and Affiliations

  1. School of Life Sciences, Shandong University, 27 Shanda South Road, Jinan, Shandong, 250100, China

    Xiu-Zhen Shi, Xiao-Fan Zhao & Jin-Xing Wang

Authors
  1. Xiu-Zhen Shi
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  2. Xiao-Fan Zhao
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  3. Jin-Xing Wang
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Correspondence to Jin-Xing Wang.

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Published in Russian in Biokhimiya, 2008, Vol. 73, No. 6, pp. 854–862.

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Shi, XZ., Zhao, XF. & Wang, JX. Molecular cloning and analysis of function of nucleoside diphosphate kinase (NDPK) from the scallop Chlamys farreri . Biochemistry Moscow 73, 686–692 (2008). https://doi.org/10.1134/S0006297908060096

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  • DOI: https://doi.org/10.1134/S0006297908060096

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