TY - JOUR
T1 - Molecular cloning and characterization of alpha 2-macroglobulin (α2-M) from the haemocytes of Chinese mitten crab Eriocheir sinensis
AU - Qin, Chuanjie
AU - Chen, Liqiao
AU - Qin, Jianguang
AU - Zhao, Daxian
AU - Zhang, Hao
AU - Wu, Ping
AU - Li, Erchao
AU - Yu, Na
PY - 2010/8
Y1 - 2010/8
N2 - Alpha 2-macroglobulin (α2-M) is a large protein in blood and is able to inactivate a variety of proteinases in invertebrates and vertebrates. The α2-M gene was obtained from the Chinese mitten crab, Eriocheir sinensis, by the reverse transcript-polymerase chain reaction (RT-PCR) and the rapid amplification of cDNA ends (RACE). The α2-M cDNA of E. sinensis contained 4851 bp with an open reading frame of 4371 bp, a 70 bp 5'-untranslated region, a 410 bp 3'-untranslated region, and three common putative functional domains. The putative domains include a bait region, a GCGEQNM thiol ester domain, and a receptor-binding domain in E. sinensis, which are similar to those in other species. Sequence comparison indicates that the α2-M deduced amino acid sequence of E. sinensis has an overall identity of 44%, 44%, 43% and 38% to that of Marsupenaeus japonicus, Macrobrachium rosenbergii, Litopenaeus vannamei and Scylla serrata, respectively. Alignment of deduced amino acid sequence to other species shows that the overall structure of α2-M is evolutionarily conserved. Phylogenetic analysis reveals that the E. sinensis α2-M is closely related to the α2-Ms in other arthropods. Quantitative PCR analysis showed that α2-M was mainly expressed in haemocytes, but not in gill, muscle, hepatopancreas, gut and stomach. Its mRNA transcript in haemocytes of E. sinensis increased significantly at 12, 24 and 48 h post-Aeromonas hydrophila (Gram negative bacteria) injection, indicating that A. hydrophila could induce α2-M mRNA expression in E. sinensis.
AB - Alpha 2-macroglobulin (α2-M) is a large protein in blood and is able to inactivate a variety of proteinases in invertebrates and vertebrates. The α2-M gene was obtained from the Chinese mitten crab, Eriocheir sinensis, by the reverse transcript-polymerase chain reaction (RT-PCR) and the rapid amplification of cDNA ends (RACE). The α2-M cDNA of E. sinensis contained 4851 bp with an open reading frame of 4371 bp, a 70 bp 5'-untranslated region, a 410 bp 3'-untranslated region, and three common putative functional domains. The putative domains include a bait region, a GCGEQNM thiol ester domain, and a receptor-binding domain in E. sinensis, which are similar to those in other species. Sequence comparison indicates that the α2-M deduced amino acid sequence of E. sinensis has an overall identity of 44%, 44%, 43% and 38% to that of Marsupenaeus japonicus, Macrobrachium rosenbergii, Litopenaeus vannamei and Scylla serrata, respectively. Alignment of deduced amino acid sequence to other species shows that the overall structure of α2-M is evolutionarily conserved. Phylogenetic analysis reveals that the E. sinensis α2-M is closely related to the α2-Ms in other arthropods. Quantitative PCR analysis showed that α2-M was mainly expressed in haemocytes, but not in gill, muscle, hepatopancreas, gut and stomach. Its mRNA transcript in haemocytes of E. sinensis increased significantly at 12, 24 and 48 h post-Aeromonas hydrophila (Gram negative bacteria) injection, indicating that A. hydrophila could induce α2-M mRNA expression in E. sinensis.
KW - Aeromonas hydrophila
KW - Alpha 2-macroglobulin
KW - Eriocheir sinensis
KW - Innate immunity
KW - Protease inhibitor
KW - Thiol ester
UR - http://www.scopus.com/inward/record.url?scp=77955698904&partnerID=8YFLogxK
U2 - 10.1016/j.fsi.2010.03.010
DO - 10.1016/j.fsi.2010.03.010
M3 - Article
VL - 29
SP - 195
EP - 203
JO - Fish and Shellfish Immunology
JF - Fish and Shellfish Immunology
SN - 1050-4648
IS - 2
ER -