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|Title: ||Molecular cloning and characterisation of a proteinase inhibitor, alpha 2-macroglobulin (α2-M) from the haemocytes of tiger shrimp Penaeus monodon|
|Authors: ||Yong-Chin Lin;Baskaralingam Vaseeharan;Chi-Fong Ko;Tzu-Ting Chiou;Jiann-Chu Chen|
|Contributors: ||NTOU:Department of Aquaculture|
|Keywords: ||Penaeus monodon;Alpha 2-macroglobulin;Protease inhibitor;Thioester-containing protein;Bait region;Innate immunity;Peptidoglycan|
|Issue Date: ||2011-10-21T02:18:27Z
|Publisher: ||Molecular Immunology|
|Abstract: ||Abstract:An alpha 2-macroglobulin ( 2-M) gene was cloned from the haemocytes of tiger shrimp Penaeus monodon by RT-PCR, cloning and sequencing of overlapping PCR and rapid amplification of cDNA ends (RACE) method. Analysis of the nucleotide sequence revealed that the 2-M cDNA consists of 4876 bp with an open reading frame (ORF) of 4494 bp, a 52 bp 5 -untranslated region, and a 327 bp 3 -untranslated region containing a poly A signal. The open reading frame encodes a protein of 1498 amino acids with 18 residues signal sequence. The predicted molecular mass of the mature protein (1480 amino acids) is 167.7 kDa with an estimated pI of 5.30. The P. monodon 2-M sequence contains putative functional domains including a GCGEQNM thioester region, a bait region, and a receptor-binding domain which are present in other invertebrate and vertebrate 2-Ms. Sequence comparison showed that 2-M deduced amino acid sequence of P. monodon has an overall similarity of 85, 52 and 49% to that of kuruma shrimp Marsupenaeus japonicus, American horseshoe crab Limulus polyphemus and mud crab Scylla serrata, respectively. Alignment of the deduced amino acid sequence to other species 2-M showed that the overall structure is evolutionarily conserved and phylogentic analysis revealed that P. monodon 2-M is closely related to other arthropod 2-M, and displays the highest similarity to M. japonicus 2-M. The 2-M was mainly expressed in haemocytes, but not in eyestalk, gill, muscle, hepatopancreas, and intestine. Quantitative real-time RT-PCR analysis showed that 2-M mRNA transcript in haemocytes of P. monodon increased significantly in 12, 24 and 48 h post-peptidoglycan (PG) injection, but returned to the original values in 72 h post-PG injection. ?2006 Elsevier Ltd. All rights reserved.|
|Relation: ||44(6), pp.1065-1074|
|Appears in Collections:||[水產養殖學系] 期刊論文|
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