TY - JOUR
T1 - A macrophage migration inhibitory factor like gene from scallop Chlamys farreri
T2 - Involvement in immune response and wound healing
AU - Li, Fengmei
AU - Huang, Shuyan
AU - Wang, Lingling
AU - Yang, Jialong
AU - Zhang, Huan
AU - Qiu, Limei
AU - Li, Ling
AU - Song, Linsheng
PY - 2011/1
Y1 - 2011/1
N2 - Macrophage migration inhibitory factor (MIF) is an evolutionarily ancient and highly conserved cytokine with multiple functions. In the present study, a MIF-like gene was cloned from Zhikong scallop Chlamys farreri (designated as CfMIF) based on expressed sequence tag (EST) analysis and rapid amplification of cDNA ends (RACE) approach. The full-length cDNA of CfMIF was of 2296bp, consisting of a 5' untranslated region (UTR) of 60bp, a 3' UTR of 1903bp with a poly(A) tail and an open reading frame (ORF) of 333bp encoded 111 amino acid residues with a calculated molecular mass of 12.6kDa and a theoretical isoelectric point of 5.63. The deduced amino acid sequence of CfMIF shared 27-50.5% similarity with those of other known MIFs. A conserved MIF domain was identified in the deduced amino acid sequence of CfMIF, and conserved proline2 and lysine33 were also found to be present in CfMIF. Phylogenetic analysis revealed that CfMIF is one of MIF members. The tissue distribution and temporal expression of CfMIF in hemocytes of scallop after lipopolysaccharide (LPS), peptidoglycan (PGN) and β-glucan stimulation were detected by real-time RT-PCR. CfMIF gene was ubiquitously expressed in six selected tissues of healthy scallops, with the higher expression levels in hepatopancreas, mantle and gill. In comparison with the control group, the expression of CfMIF mRNA in hemocytes was up-regulated significantly at 6h, 24h and 48h after LPS treatment, and at all time points after PGN and glucan treatment. The cDNA fragment encoding mature peptide of CfMIF was recombined and expressed in Escherichia coli BL21 (DE3) pLysS. The recombinant protein of CfMIF (rCfMIF) promoted sheep fibroblast migration into scraped spaces in vitro. These results generated from the present study encourage us to suggest that CfMIF was a novel member of MIF family, and it was involved in immune response and wound healing by promoting fibroblast migration.
AB - Macrophage migration inhibitory factor (MIF) is an evolutionarily ancient and highly conserved cytokine with multiple functions. In the present study, a MIF-like gene was cloned from Zhikong scallop Chlamys farreri (designated as CfMIF) based on expressed sequence tag (EST) analysis and rapid amplification of cDNA ends (RACE) approach. The full-length cDNA of CfMIF was of 2296bp, consisting of a 5' untranslated region (UTR) of 60bp, a 3' UTR of 1903bp with a poly(A) tail and an open reading frame (ORF) of 333bp encoded 111 amino acid residues with a calculated molecular mass of 12.6kDa and a theoretical isoelectric point of 5.63. The deduced amino acid sequence of CfMIF shared 27-50.5% similarity with those of other known MIFs. A conserved MIF domain was identified in the deduced amino acid sequence of CfMIF, and conserved proline2 and lysine33 were also found to be present in CfMIF. Phylogenetic analysis revealed that CfMIF is one of MIF members. The tissue distribution and temporal expression of CfMIF in hemocytes of scallop after lipopolysaccharide (LPS), peptidoglycan (PGN) and β-glucan stimulation were detected by real-time RT-PCR. CfMIF gene was ubiquitously expressed in six selected tissues of healthy scallops, with the higher expression levels in hepatopancreas, mantle and gill. In comparison with the control group, the expression of CfMIF mRNA in hemocytes was up-regulated significantly at 6h, 24h and 48h after LPS treatment, and at all time points after PGN and glucan treatment. The cDNA fragment encoding mature peptide of CfMIF was recombined and expressed in Escherichia coli BL21 (DE3) pLysS. The recombinant protein of CfMIF (rCfMIF) promoted sheep fibroblast migration into scraped spaces in vitro. These results generated from the present study encourage us to suggest that CfMIF was a novel member of MIF family, and it was involved in immune response and wound healing by promoting fibroblast migration.
KW - Chlamys farreri
KW - Fibroblast migration
KW - Immune response
KW - MRNA expression
KW - Macrophage migration inhibitory factor
UR - https://www.scopus.com/pages/publications/77957862827
U2 - 10.1016/j.dci.2010.08.009
DO - 10.1016/j.dci.2010.08.009
M3 - 文章
C2 - 20804783
AN - SCOPUS:77957862827
SN - 0145-305X
VL - 35
SP - 62
EP - 71
JO - Developmental and Comparative Immunology
JF - Developmental and Comparative Immunology
IS - 1
ER -