A transglutaminase from Chinese shrimp (Fenneropenaeus chinensis), full-length cDNA cloning, tissue localization and expression profile after challenge.
Transglutaminase can catalyze the cross-linking reaction between soluble clotting protein molecules from the plasma for prevention of excess blood loss from a wound and obstructing micro-organisms from invading the wound in crustaceans. A novel transglutaminase (FcTG) gene was cloned from hemocytes of Chinese shrimp Fenneropenaeus chinensis by 3' and 5' rapid amplification of cDNA ends (RACE) PCR. The full-length cDNA consists of 2972bp, encoding 757 amino acids with a calculated molecular mass of 84.96kDa and a theoretical isoelectric point of 5.61. FcTG contains a typical transglutaminase-like homologue (TGc domain: E-value=1.94e-38). Three catalytic sites (Cys-324, His-391 and Asp-414) are present in this domain. The deduced amino acid sequence of FcTG showed high identity with black tiger shrimp TG, kuruma shrimp TG and crayfish TG. Transcripts of FcTG mRNA were mainly detected in gill, lymphoid organ and hemocytes by RT-PCR. RNA in situ hybridization further confirmed that FcTG was constitutively expressed in hemocytes both in the circulatory system and lymphoid organ. The variation of mRNA transcription level in hemocytes and lymphoid organ following injection of killed bacteria or infection with white spot syndrome virus (WSSV) was quantified by RT-PCR. The up-regulated expression of FcTG in shrimp lymphoid organ following injection of bacteria indicates that it is inducible and might be associated with bacterial challenge.
Liu
Yi-Chen
YC
Institute of Oceanology, Chinese Academy of Sciences, No. 7 Nanhai Road, Qingdao, Shandong 266071, China.
Li
Fu-Hua
FH
Wang
Bing
B
Dong
Bo
B
Zhang
Qing-Li
QL
Luan
Wei
W
Zhang
Xiao-Jun
XJ
Xiang
Jian-Hai
JH
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D017931:DNA Primers / Q000737:chemistry; D020869:Gene Expression Profiling / Q000662:veterinary; D015971:Gene Expression Regulation, Enzymologic / Q000276:immunology*; D006434:Hemocytes / Q000502:physiology; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D033561:Penaeidae / Q000201:enzymology* / Q000235:genetics / Q000382:microbiology / Q000821:virology; D010802:Phylogeny; D016415:Sequence Alignment; D013211:Staphylococcus aureus / Q000472:pathogenicity; D013997:Time Factors; D011503:Transglutaminases / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics*; D014733:Vibrio / Q000472:pathogenicity; D046848:White spot syndrome virus 1 / Q000472:pathogenicity
D017931:DNA Primers; D011503:Transglutaminases