7,468 research outputs found
Sequence and the expression construct of SP-LL-37.
No full text(A) Nucleotide sequence of SP-LL-37 containing the coding region and stop codon as well as the signal peptide (underlined). (B) Schematic diagram of the expression construct pSP1::SP-LL-37. Detection of Nos gene copy numbers in the T0 generation of pPZP::SP-LL-37 plants. (C) PCR analysis of SP-LL-37 transgenic rice. Lanes: M; molecular marker, P; pPZP::SP-LL-37 vector, NT; non-transformed (NT) control, 1~25; independent T0 transgenic lines. (D) TaqMan PCR analysis for copy number assays using TaqMan probe for single copy selection in T0 transgenic Rice. +; single copy, -; multi copy, T2-homozygous and T2-heterozygous; single copy control, NT; negative control, WT; wild type control.</p
Identification of pPZP::SP-LL-37 insertion transgenic plants.
No full textThe genomic structures of insertion alleles were determined by FST analysis in which boxes, bold lines, and triangles indicate exons, intron, and pPZP::SP-LL-37, respectively. The arrow and arrowhead indicate gene specific primer pairs from genomic DNA. Genomic DNA was isolated from the leaves of pPZP::SP-LL-37 regenerated plants for PCR analysis. WT: wild type, T: T0 plant with single T-DNA insert for pPZP::SP-LL-37 insertion line.</p
Subcellular localization of the SP-LL-37 protein in tobacco.
No full textGFP protein was attached to the end of C-terminal of LL-37 protein to see the localization in tobacco cells. (A) Cartoon image of constructs used in subcellular localization. (B) Fluorescence image of cell membrane (N. benthamiana epidermal cells) from Agrobacterium-mediated transient expression (using the p19 protein to enhance the expression level) and cytoplasm expressing the SP-LL-37 protein with positive and negative control by confocal microscopy.</p
Generation and molecular analysis of the transgenic lines expressing pPZP::SP-LL-37.
No full text(A) TaqMan PCR analysis for copy number assays using TaqMan probe for selected homozygous T1 plants; T2 homo; T2-homozygous, T2 hetero; T2-heterozygous, NT; no template, WT; wild type, 6–1~20–3; 19 T1 plants. (B) SP-LL-37 gene expression in T1 homo transgenic lines using RT-PCR. Total RNA was isolated from each plant, and 0.5 μg of this RNA was amplified with SP-LL-37-specific primers. Rice actin gene was amplified as a loading control. (C) ELISA analysis of SP-LL-37 lines in bovine sperm conditioned media collected at T1-transgenic rice plants.</p
Copy number frequency of pPZP::SP-LL-37 in transgenic plants.
No full textCopy number frequency of pPZP::SP-LL-37 in transgenic plants.</p
Agronomic traits of pPZP:: SP-LL-37 T<sub>1</sub> homo lines.
No full textAgronomic traits of pPZP:: SP-LL-37 T1 homo lines.</p
The modulatory effect of TLR2 on LL-37-induced human mast cells activation
No full textThe sole and endogenous anti-microbial peptide LL-37 is a significant effector molecule in the innate host defense system. Apart from its broadly direct anti-microbial activity, the peptide also activates mast cell in respect of allergic diseases and inflammation. On the other hand, mast cell can be activated by Toll-like receptors (TLRs) which are at the center of innate immunity. It was the aim of the study to illustrate the modulatory effect of TLR2 ligands peptidoglycan (PGN) and tripalmitoyl-S-glycero-Cys-(Lys)4 (Pam3CSK4) on LL-37 induced LAD2 cells (a human mast cell line) activation. LL-37 induced LAD2 cells degranulation and the release of IL-8. TLR2 ligands didn't induce LAD2 cells degranulation, but triggered the release of IL-8. Incubation with PGN or Pam3CSK4 significantly suppressed LL-37-induced degranulation through inhibition of calcium mobilization from LAD2 cells. Similarly, the release of IL-8 was inhibited when LAD2 cells were co-stimulated with TLR2 ligands and LL-37. Studies with inhibitors of key enzymes involved in mast cell signaling indicated that the release of IL-8 induced by TLR2 ligands and LL-37 involved the activation of the PI3K, ERK, JNK and calcineurin signaling pathways. In contrast, p38 activation down-regulated the release of IL-8 induced by TLR2 ligands and LL-37. Taken together, these observations suggest that activation of human mast cells by LL-37 could be modified by TLR2 ligands and the function of human mast cells could be switched from allergic reactions to innate immune response. (C) 2016 Elsevier Inc. All rights reserved.National Natural Science Foundation of China [81271755, 81371737]; Guangdong Natural Science Foundation [2014A030313708]; Shenzhen Research Grant [CXZZ20140416144209739, JCYJ20130329110752142, KQCX20120803145850990]SCI(E)[email protected]; [email protected]
Flanking T-DNA analysis from pPZP::SP-LL-37 T<sub>0</sub> plants.
No full textFlanking T-DNA analysis from pPZP::SP-LL-37 T0 plants.</p
SP-LL-37, human antimicrobial peptide, enhances disease resistance in transgenic rice.
No full textHuman LL-37 is a multifunctional antimicrobial peptide of cathelicidin family. It has been shown in recent studies that it can serve as a host's defense against influenza A virus. We now demonstrate in this study how signal peptide LL-37 (SP-LL-37) can be used in rice resistance against bacterial leaf blight and blast. We synthesized LL-37 peptide and subcloned in a recombinant pPZP vector with pGD1 as promoter. SP-LL-37 was introduced into rice plants by Agrobacterium mediated transformation. Stable expression of SP-LL-37 in transgenic rice plants was confirmed by RT-PCR and ELISA analyses. Subcellular localization of SP-LL-37-GFP fusion protein showed evidently in intercellular space. Our data on testing for resistance to bacterial leaf blight and blast revealed that the transgenic lines are highly resistant compared to its wildtype. Our results suggest that LL-37 can be further explored to improve wide-spectrum resistance to biotic stress in rice
LL-37-induced human mast cell activation through G protein-coupled receptor MrgX2
No full textHuman LL-37 is an important class of cationic antimicrobial peptide (CAP) that is known to stimulate mast cell activation. While many studies have been conducted on LL-37, to date little is known about the functional receptors for LL-37-induced human mast cell activation, in particular in terms of the release of de novo synthesized mediators. Thus, the aim of the present study is to identify the functional receptors for LL-37-induced human mast cell activation in terms of the degranulation and release of de novo synthesized mediators and investigate the downstream signalling pathways involved in mast cell activation. Overall, our study importantly demonstrates that LL-37-induced human mast cell degranulation and release of de novo synthesized mediators function primarily through the activation of MrgX2. We furthermore show that LL-37-induced human mast cell line LAD2 cells are involved in the degranulation and release of IL-8, and that FPRL1 and P2X7 have only a partial effect on IL-8 release, and no effect on mast cell degranulation triggered by LL-37. Instead, we find that silencing the expression of MrgX2 in human mast cell significantly inhibits the LL-37-induced degranulation and release of IL-8. Overall, this effect is associated with the activation of the Gi protein, PLC/PKC/Calcium/NFAT, PI3K/Akt and MAPKs signalling pathways.National Natural Science Foundation of China [81401299]; Natural Science Foundation of Guangdong Province [2014A030313711]; Characteristic innovation project of Guangdong Province Ordinary University [2015KTSCX120]; Shenzhen Peacock Plan [827-000209]SCI(E)ARTICLE6-124
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