Molecular characterization of trypsinogens and development of trypsinogen gene expression and tryptic activities in grass carp (Ctenopharyngodon idellus) and topmouth culter (Culter alburnus).
This study examined the gene structures and expression of trypsinogens, as well as the trypsin activities of the grass carp Ctenopharyngodon idellus (herbivorous) and the topmouth culter Culter alburnus (carnivorous), which are commercially important freshwater species of the family Cyprinidae in China. Isolated full-length trypsinogen cDNA clones were 869 bp and 857 bp. The deduced amino acid sequences were 242aa and 247aa long, both containing the highly conserved residues essential for serine protease catalytic and conformational maintenance. The results from isoelectric and phylogenetic analyses suggest that grass carp trypsinogen is grouped with teleost trypsinogen group I, while topmouth culter trypsinogen is grouped with group II. The expression pattern of trypsinogen mRNA was similar between these two species, appearing 2 days post-hatching (dph) and reaching peaks at 11 and 23 dph. The trypsin-specific activities in both species were detected 2 dph and reached the major peaks at 8 dph, however the minor peaks were observed at 20 dph in the grass carp and 17 dph in the topmouth culter. The trypsin-specific activity was significantly higher in the grass carp than in the topmouth culter, which may be attributed to the nature of their different nutritional habits.
College of Fisheries, Key Lab of Freshwater Biodiversity Conservation and Utilization, Ministry of Agriculture, Huazhong Agricultural University, Wuhan 430070, PR China.
Guo-Liang
GL
Ruan
Yang
Y
Li
Ze-Xia
ZX
Gao
Huan-Ling
HL
Wang
Wei-Min
WM
Wang
D000595:Amino Acid Sequence; D000818:Animals; D002347:Carps / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D003001:Cloning, Molecular; D003530:Cyprinidae / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D029941:Fish Proteins / Q000235:genetics* / Q000378:metabolism; D020869:Gene Expression Profiling; D018507:Gene Expression Regulation, Developmental; D015971:Gene Expression Regulation, Enzymologic; D007814:Larva / Q000201:enzymology / Q000235:genetics / Q000254:growth & development; D008969:Molecular Sequence Data; D010802:Phylogeny; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D014357:Trypsin / Q000378:metabolism; D014362:Trypsinogen / Q000145:classification / Q000235:genetics* / Q000378:metabolism
D018076:DNA, Complementary; D029941:Fish Proteins; D014362:Trypsinogen; D014357:Trypsin