Biochemistry (Moscow)

, Volume 73, Issue 6, pp 686–692 | Cite as

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.

Key words

scallop Chlamys farreri nucleoside diphosphate kinase (NDPK) expression profile recombinant expression 

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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Copyright information

© MAIK Nauka 2008

Authors and Affiliations

  1. 1.School of Life SciencesShandong UniversityJinanChina

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