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Molecular cloning and expression analysis of crustin-like gene from Chinese shrimp Fenneropenaeus chinensis

Acta Oceanologica Sinica 2008年第2期27卷 81-92页

作者： LIU Fengsong LI Fuhua XIANG Jianhai DONG Bo LIU Yichen ZHANG Xiaojun ZHANG Liusuo College of Life Science Hebei University Baoding 071002 China Experimental Marine Biology Laboratory Institute of Oceanology Chinese Academy of Sciences Qingdao 266071 China

A new member of antimicrobial protein genes of the crustin family was cloned from haemocytes of the Chinese shrimp Fennero- penaeus chinensis by 3 ′and 5′ RACE. The full-length cDNA of crustin-like gene contains a 390 bp open reading frame, encoding 130 amino acids. The deduced peptide contains a putative signal peptide of 17 amino acids and mature peptide of 113 amino acids. The molecular mass of the deduced mature peptide is 12. 3 ku. It is highly cationic with a theoretical isoelectric point of 8.5. The deduced amino acids sequence of this crustin showed high homology with those of Penaeus （ Litopenaeus ） setferus. Northern blotting showed that the cloned crustin gene was mainly expressed in haemocytes, gill, intestine, and RNA in situ hybridization indicated that the crustin gene was constitutively expressed exclusively in haemocytes of these tissues. Capillary elec- trophoresis RT-PCR analysis showed that crustin was up-regulated dramatically from 12 to 48 h after a brief decrease of mRNA during first 6 h in response to microbe infection. The level of crustin mRNA began to restore at 72 h post-challenge. This indicated that crustin gene might play an important role when shrimps are infected by bacterial pathogen.

关键词： crustin-like gene Fenneropenaeius chinensis molecular cloning expression analysis microbe challenge

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