Rojtinnakorn Jiraporn, Hirono Ikuo, Itami Toshiaki, Takahashi Yukinori, Aoki Takashi
Department of Aquatic Biosciences, Tokyo University of Fisheries, Japan.
Fish Shellfish Immunol. 2002 Jul;13(1):69-83. doi: 10.1006/fsim.2001.0382.
Gene expression in haemocytes of the kuruma prawn (Penaeus japonicus) was investigated using an expressed sequence tag (EST) approach. Partial nucleotide sequences of cDNA library clones constructed from normal and white spot syndrome virus (WSSV)--infected P. japonicus haemocytes were determined. Of 635 clones obtained from the normal library, 284 (44.7%) significantly matched sequences in GenBank, and of 370 clones obtained from WSSV-infected library, 174 (47.0%) significantly matched sequences in the database. One hundred fifty-two deduced proteins were newly identified. Of these, 28 types were involved in biodefence. For the prophenoloxidase system, there are prophenoloxidase, coagulation factor G-beta chain precursor, factor D, Masquarade-like protease, transglutaminase (TGase), clottable protein and eight types of protease inhibitors (two types of antileukoproteinase, alpha-2-macroglobulin, chelonianin, elastase inhibitor, two types of Kazal inhibitor and Kunitz-type inhibitor). For antibacterial peptides, there are bactinecin 11, penaeidin-2 precursor and lysozyme c type. The others defence-related proteins are basophil leukocyte interleukin-3-regulated protein, natural killer enhancing factor (NK-EF), integral membrane protein (CD34+), ESM-1, Notch homologue and Drac homologue. For the adhesion proteins, there are beta-integrin, cell adhesion molecule (CAM) and three types of collagens. All ESTs representing protease inhibitors and tumour-related proteins were found only in the WSSV-infected library. Those encoding for apoptotic peptides were expressed at high levels in infected library. The putative defence proteins accounted for 2.7% of total ESTs in a normal shrimp library and 15.7% of the total ESTs in an infected library.
采用表达序列标签(EST)方法研究了日本对虾血细胞中的基因表达。测定了从正常和感染白斑综合征病毒(WSSV)的日本对虾血细胞构建的cDNA文库克隆的部分核苷酸序列。从正常文库获得的635个克隆中,284个(44.7%)与GenBank中的序列有显著匹配;从感染WSSV的文库获得的370个克隆中,174个(47.0%)与数据库中的序列有显著匹配。新鉴定出152种推导蛋白。其中,28种参与生物防御。对于酚氧化酶原系统,有酚氧化酶原、凝血因子G-β链前体、因子D、类假面舞会蛋白酶、转谷氨酰胺酶(TGase)、可凝蛋白和8种蛋白酶抑制剂(2种抗白细胞蛋白酶、α-2-巨球蛋白、海龟蛋白、弹性蛋白酶抑制剂、2种Kazal抑制剂和Kunitz型抑制剂)。对于抗菌肽,有杆菌素11、对虾抗菌肽-2前体和溶菌酶c型。其他防御相关蛋白有嗜碱性白细胞白细胞介素-3调节蛋白、自然杀伤增强因子(NK-EF)、整合膜蛋白(CD34+)、ESM-1、Notch同源物和Drac同源物。对于粘附蛋白,有β-整合素、细胞粘附分子(CAM)和3种胶原蛋白。所有代表蛋白酶抑制剂和肿瘤相关蛋白的EST仅在感染WSSV的文库中发现。编码凋亡肽的EST在感染文库中高表达。推测的防御蛋白在正常对虾文库中占总EST的2.7%,在感染文库中占总EST的15.7%。