Molecular cloning and characterization of macrophage migration inhibitory factor from small abalone Haliotis diversicolor supertexta.
ABSTRACT The macrophage migration inhibitory factor (mif) cDNA and its genome were cloned from small abalone Haliotis diversicolor supertexta. Small abalone mif (samif) was originally identified from an expressed sequence tag (EST) fragment from a normalized cDNA library. It's 5' untranslated region (UTR) was obtained by 5' rapid amplification of cDNA end (RACE) techniques and its genomic DNA was cloned by PCR. The full-length cDNA of samif was of 535 bp, consisting of a 5'-terminal UTR of 49 bp, an open reading frame of 384 bp and a 3'-terminal UTR of 102 bp. The deduced protein was composed of 128 amino acids, with an estimated molecular mass of 14.0 kDa and a predicted pI of 6.90. The full-length samif genomic DNA comprises 3238 bp, containing three exons and two introns. Real time quantitative PCR analysis revealed that samif gene is constitutively expressed in 6 selected tissues, and its expression level in hepatopancreas is higher than that in the other tissues (p < 0.01). Samif expression level in the hepatopancreas at 24 and 48 h after Vibrio parahaemolyticus injection was upregulated significantly (p < 0.01), but there was no significant change after exposure to tributyltin (TBT) (p > 0.05).
- SourceAvailable from: Frank Melzner[Show abstract] [Hide abstract]
ABSTRACT: The marine mussel Mytilus edulis and its closely related sister species are distributed world-wide and play an important role in coastal ecology and economy. The diversification in different species and their hybrids, broad ecological distribution, as well as the filter feeding mode of life has made this genus an attractive model to investigate physiological and molecular adaptations and responses to various biotic and abiotic environmental factors. In the present study we investigated the immune system of Mytilus, which may contribute to the ecological plasticity of this species. We generated a large Mytilus transcriptome database from different tissues of immune challenged and stress treated individuals from the Baltic Sea using 454 pyrosequencing. Phylogenetic comparison of orthologous groups of 23 species demonstrated the basal position of lophotrochozoans within protostomes. The investigation of immune related transcripts revealed a complex repertoire of innate recognition receptors and downstream pathway members including transcripts for 27 toll-like receptors and 524 C1q domain containing transcripts. NOD-like receptors on the other hand were absent. We also found evidence for sophisticated TNF, autophagy and apoptosis systems as well as for cytokines. Gill tissue and hemocytes showed highest expression of putative immune related contigs and are promising tissues for further functional studies. Our results partly contrast with findings of a less complex immune repertoire in ecdysozoan and other lophotrochozoan protostomes. We show that bivalves are interesting candidates to investigate the evolution of the immune system from basal metazoans to deuterostomes and protostomes and provide a basis for future molecular work directed to immune system functioning in Mytilus.PLoS ONE 01/2012; 7(3):e33091. · 3.73 Impact Factor
- [Show abstract] [Hide abstract]
ABSTRACT: A polysaccharide Astragalus polysaccharide (APS) was obtained from the boiling water extract of Astragalus membranaceus (Fisch.) Bge. var. mongholicus (Bge.) Hsiao. The results of gas chromatography (GC) indicated that APS consisted of l-rhamnose (l-Rha), d-xylose (d-Xyl), d-glucose (d-Glc), and d-galactose (d-Gal) in the molar ratio of 1:4:5:1.5. Its molecular weight was determined to be 3.01 × 10(5) by high-performance gel filtration chromatography. The results of (1)H NMR and (13)C NMR spectral analysis indicated that APS had a linear backbone mainly consisting of 1,3-linked β-d-Gal residues with insertion of β-Glc, 1,6-linked α-Gal, 1,5-linked β-Xyl, 1,4-linked β-Gal, β-d-Gal, 1,2-linked α-Rha, 1,2,4-linked α-Rha residues. HSQC spectrum indicated that C-2 and C-6 may link with H.Journal of Asian natural products research 05/2013; · 0.61 Impact Factor
- [Show abstract] [Hide abstract]
ABSTRACT: 544 INTRODUCTION Macrophage migration inhibitory factor (MIF) is a mysterious cytokine and is reportedly released by antii gennactivated T lymphocytes with the function of inhibiting vertebrate macrophage migration [1, 2]. Since it was found in 1966, more studies on MIF have been undertaken, showing that it is produced not only by activated T cells but by other immune cells such as monocytes, macrophages, and some nonnimmune cells [3, 4]. MIFs have been isolated and characterized from many organisms like the bovine brain [5, 6] and rat epididymis . Since human MIF complementary cDNA was first cloned in 1989, many MIF genes have been cloned successfully from various species such as Mus musculus [8, 9], the Qingdao amphioxus Brann chiostoma belcheri tsingtaunese , and the sheep Ovis aries . Recently, MIF has been reported to play many important roles in the internal environn ment. It has been confirmed to act as a mediator durr ing the inflammatory response [12, 13, 14]. Many other studies have shown that MIF is also involved in the processes of regulating cell proliferation and tumor angiogenesis . Ito et al demonstrated that MIF also functions as a growth factor during the period of embryonic differentiation . In addition, Yan et al  suggested that MIF produced at superrphysiological levels in a murine neuroblastoma cell line (Neuroo2a) inhibited TTcell activation and induced the death of TT cells through an γIFN pathway. Besides, studies [4, 17, 18] have also implied that MIF could activate the ERK1–ERK2–MAPK pathway of the intracellular signal system. Oxana et al  suggested that at heat stress temperatures, when heat shock proteins are not abundant, MIF may act as a chaperone to bind other proteins of similar properties to counteract the early onset of stress. Mud crabs (Scylla) are mostly distributed throughh out the IndooPacific and Indian Ocean regions  as one of the most commercially valuable species in these areas. However huge economic losses are caused each year by the death of mud crabs from diseases associi ated with Vibrio parahaemolyticus, the most important infectious agent. To enhance the immunity of mud crabs, researchers have paid more attention to the mechanisms of action of immunogenic proteins. Although the cloning of MIF from many species has been reported, there is no report about the cloning of MIF from Scylla paramamosain (SpMIF). In this study, we characterized the fullllength cDNA of