Cloning and characterization of hemolymph clottable proteins of kuruma prawn (Marsupenaeus japonicus) and white shrimp (Litopenaeus vannamei)

Winton Cheng, Inn H. Tsai, Chang J. Huang, Pei C. Chiang, Chia Hsiung Cheng, Maw Sheng Yeh

研究成果: 雜誌貢獻文章同行評審

26 引文 斯高帕斯(Scopus)

摘要

The hemolymph clottable protein (CP) of Marsupenaeus japonica (designated as Mj-CP) was purified by a DEAE anion-exchanger and a Sepharose CL-6B gel filtration column. In the presence of Ca2+, it formed stable clots in vitro upon the addition of the hemocytes lysate containing transglutaminase. Results of gel filtration chromatography and SDS-PAGE indicated that Mj-CP mainly existed as disulfide-linked homodimers of 390 kDa. Specific primers were designed; PCR as well as RACE help to clone and sequence Mj-CP cDNA of 5660 bp. The predicted CP-precursor contains a signal peptide followed by a subunit of 1671 amino acids (isoelectric point 5.6), including two RGD motifs and three potential N-glycosylation sites. Mj-CP is structurally 80% and 38% identical to the CPs of tiger shrimp and crayfish, respectively. Likewise, CP cDNA of white shrimp (Litopenaeus vannamei) was also cloned and sequenced; the predicted CP has 1666 amino acid residues and an isoelectric point of 5.2. Both CPs bear potential transglutaminase cross-linking sites, i.e. seven Ser-Lys-Thr repeats near the N-terminus, a Ser- and Gln-rich region in the middle, and polyGln (n=8-11) near the C-terminus. Phylogenetic analyses of crustacean CPs and vitellogenins revealed divergent evolution of the two protein families. By RT-PCR, the sub-cuticular epidermis was identified as one of the major tissues that express CP in M. japonica.

原文英語
頁(從 - 到)265-274
頁數10
期刊Developmental and Comparative Immunology
32
發行號3
DOIs
出版狀態已發佈 - 2008
對外發佈

ASJC Scopus subject areas

  • 發展生物學
  • 免疫學

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