Molecular cloning and characterisation of peroxinectin, a cell adhesion molecule, from the giant freshwater prawn Macrobrachium rosenbergii.
Expression of peroxinectin cDNA was determined from haemocytes of giant freshwater prawn Macrobrachium rosenbergii using oligonucleotide primers and reverse transcription polymerase chain reaction (RT-PCR) based on the peroxinectin sequence of white shrimp Litopenaeus vannamei, tiger shrimp Penaeus monodon, and freshwater crayfish Pacifastacus leniusculus. The peroxinectin of M. rosenbergii was constitutively expressed. Analysis of the nucleotide sequence revealed that the cDNA clone has an open reading frame of 2,403 bp encoding a protein of 801 amino acids including a 20 amino acid signal peptide. The calculated molecular mass of the mature protein (781 amino acids) was 88.7 kDa with an estimated pI of 6.8. A putative peroxidase domain and a putative integrin-binding motif, KGD (Lys-Gly-Asp) were observed in prawn peroxinectin at the C-terminal. Sequence comparison showed that peroxinectin deduced amino acid of M. rosenbergii had an overall similarity of 62%, 64%, and 66% to that of P. leniusculus, P. monodon, and L. vannamei, respectively. Quantitative real-time PCR analysis showed that peroxinectin transcript in haemocyte of M. rosenbergii decreased significantly after 3, 6 and 12h injection with Lactococcus garvieae.
Hsu
Pei-I
PI
Department of Aquaculture, National Pingtung University of Science and Technology, 1 Hseuh-Fu Road, Nei-Pu Hsiang, Pingtung 91201, Taiwan.
Liu
Chun-Hung
CH
Tseng
Deng-Yu
DY
Lee
Pai-Po
PP
Cheng
Winton
W
D000199:Actins / Q000032:analysis; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence / Q000235:genetics; D001798:Blood Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics* / Q000276:immunology*; D002244:Carbon / Q000008:administration & dosage; D015815:Cell Adhesion Molecules / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics* / Q000276:immunology*; D003001:Cloning, Molecular / Q000379:methods; D017931:DNA Primers / Q000737:chemistry; D018076:DNA, Complementary / Q000737:chemistry; D006434:Hemocytes / Q000737:chemistry; D043143:Hepatopancreas / Q000737:chemistry; D017045:Lactococcus / Q000276:immunology; D008969:Molecular Sequence Data; D009132:Muscles / Q000737:chemistry; D033541:Palaemonidae / Q000235:genetics* / Q000276:immunology* / Q000382:microbiology / Q000502:physiology; D012333:RNA, Messenger / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary; D016415:Sequence Alignment / Q000662:veterinary; D013997:Time Factors
D000199:Actins; D001798:Blood Proteins; D015815:Cell Adhesion Molecules; D017931:DNA Primers; D018076:DNA, Complementary; D012333:RNA, Messenger; C511086:peroxinectin protein, Macrobrachium rosenbergii; C096867:peroxinectin protein, Pacifastacus leniusculus; D002244:Carbon