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PMID:23732292
| Citation |
Li, C, Chen, S, Song, J, Liu, H, Gu, W, Ai, H, Zhao, B and Zhang, S (2013) Molecular cloning and characterization of TNFSF14 (LIGHT) and its receptor TNFRSF14 (HVEM) in guinea pig (Cavia porcellus). Gene 526:374-84 |
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| Abstract |
LIGHT (lymphotoxin-related inducible ligand that competes with herpes simplex virus (HSV) glycoprotein D for herpesvirus entry mediator on T cells) is a member of the tumor necrosis factor (TNF) ligand superfamily, which plays important roles in inflammatory and immune responses. In the present study, the cDNAs of guinea pig (Cavia porcellus) LIGHT (designated as gpLIGHT) and its receptor herpes virus entry mediator (designated as gpHVEM) were amplified from spleen by reverse transcription polymerase chain reaction (RT-PCR). The ORFs of gpLIGHT and gpHVEM cover 726 and 861 bp, encoding predicted proteins with 241 and 286 aas, respectively. The three-dimensional (3D) structure, phylogenetic relationships, and characterization of both genes were also analyzed. We also generated a 3D model to verify interaction between the two proteins. Real-time quantitative PCR (qPCR) analysis revealed that both LIGHT and HVEM are constitutively expressed in guinea pig various tissues. A fusion protein SUMO (Small Ubiquitin-like Modifier)-gpsLIGHT (the soluble mature part of gpLIGHT) was efficiently expressed in Escherichia coli BL21 (DE3) and purified using metal chelate affinity chromatography (Ni-NTA). Laser scanning confocal microscopy (LSCM) showed that gpsLIGHT can bind its receptors on T cells. The LIGHT-HVEM signaling pathway plays an important role in the immune system, and our results might provide a platform for further research into the effects of LIGHT and HVEM. |
| Links |
PubMed Online version:10.1016/j.gene.2013.05.031 |
| Keywords |
Amino Acid Sequence; Animals; Base Sequence; Binding Sites; Cell Line; Cloning, Molecular; Gene Expression; Gene Order; Guinea Pigs; Humans; Mice; Models, Molecular; Molecular Sequence Data; Organ Specificity/genetics; Phylogeny; Protein Binding; Protein Conformation; Receptors, Tumor Necrosis Factor, Member 14/chemistry; Receptors, Tumor Necrosis Factor, Member 14/genetics; Sequence Alignment; T-Lymphocytes/metabolism; Tumor Necrosis Factor Ligand Superfamily Member 14/chemistry; Tumor Necrosis Factor Ligand Superfamily Member 14/genetics; Tumor Necrosis Factor Ligand Superfamily Member 14/isolation & purification |
| edit table |
Significance
Annotations
| Gene product | Qualifier | GO Term | Evidence Code | with/from | Aspect | Extension | Notes | Status |
|---|---|---|---|---|---|---|---|---|
| GO:2000307: regulation of tumor necrosis factor (ligand) superfamily member 11 production |
ECO:0000255: |
UniProtKB:H0VCS5
|
P |
Figure 2A: nucleotide and deduced amino acid sequences of gpHVEM gene. |
complete | |||
| GO:2000307: regulation of tumor necrosis factor (ligand) superfamily member 11 production |
ECO:0000247: |
UniProtKB:H0VCS5
|
P |
Figure 2C: Amino acid sequence of HVEM for guinea pig, human, mouse, and dog. |
complete | |||
See also
References
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