服务热线: 0411-84608045 13940954745
点评详情
发布于:2019-5-17 14:34:27  访问:29 次 回复:0 篇
版主管理 | 推荐 | 删除 | 删除并扣分
, with more parasite staining at the nuclear periphery standard of an
PF11_0505 is actually a tiny RG7834MedChemExpress protein (89 amino acids) terminating in a second predicted TM; thus GFP adds a large further domain that could possibly strongly have an effect on the export efficiency from the chimeric protein. This involves two new kinds of PNEPs not present inside the query set: one with two internal hydrophobic sequences (PF11_0505) and a single using a classical Nterminal signal peptide (PF13_0194).genes from the loci of new PNEPs discovered in this study and five additional genes picked arbitrarily from other loci. All chosen candidates have been of unknown function (in line with PlasmoDB annotation), that is common for many exported proteins [6]. The selected candidate proteins had been C-terminally tagged with GFP and expressed in P. falciparum. Three GFP fusion proteins (PF07_0010-GFP, PF14_0024-GFP and PFC1035w-GFP) couldn‘t be positioned resulting from poor fluorescence. Two, PF14_0044-GFP and PF14_0046-GFP, had been not exported and showed a fluorescence pattern at the parasite periphery common of a PPM, PV or PVM location (Figure 2B). The remaining five candidates have been exported and for that reason represent new PNEPs. PF08_0004-GFP, PFF0090w-GFP, PFL0065w-GFP and PFL2515c-GFP showed a punctate pattern inside the host cell (Figure 2C) that was confirmed to represent Maurer‘s clefts (Figure S5). Cells of all of those parasite lines also showed staining in the parasite periphery and in some situations a perinuclear fluorescence indicative of an ER location, as well as the export. That is probably because of the tagging with GFP but may well also indicate a true dual location for a number of these proteins. The fifth exported protein, PF08_0005-GFP, was discovered soluble inside the host cell (once more which includes fluorescence in the meals vacuole probably from re-internalized PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/21795619 protein) as evident in the fluorescence pattern and from Western blots with selectively lysed infected RBCs (Figure 2C and D)., with further parasite staining at the nuclear periphery common of an ER place. PF11_0505-GFP also showed prominent parasite-internal fluorescence in addition to a place at the Maurer‘s clefts. PF11_0505 is often a smaller protein (89 amino acids) terminating in a second predicted TM; thus GFP adds a large further domain that may strongly affect the export efficiency on the chimeric protein. We therefore also generated a parasite line expressing a myc-tagged version of this protein that confirmed the Maurer‘s clefts place (Figure S3). In the remaining two putative exported proteins PF13_0194GFP was discovered soluble within the host cell determined by the fluorescence pattern and Western blot analysis with selectively lysed infected RBCs (Figure 1C and D). GFP fluorescence was also present within the food vacuole, most likely representing protein re-internalized in the host cell cytosol. Some cells also showed some accumulation of your protein at the parasite periphery along with the exported fraction (not shown). PF14_0045-GFP showed mobile foci at the parasite periphery and in 26 (+/25 ) of cells 1 or far more foci inside the host cell (Figure 1C, and Figure S4A and B; Video S1 and S2). The foci in the parasite periphery may possibly also represent exported protein. This interpretation is supported by two findings: firstly, Bodipy-TR-C5-ceramide staining of parasite membranes indicated that the GFP fluorescence was in close proximity to, but outside of, the parasite periphery (Figure S4A and B); secondly, pre-embedding immuno-EM resulted in a labeling of electron dense locations what appears to PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/22161446 be on the outdoors of your PVM (Figure S4C and D).
共0篇回复 每页10篇 页次:1/1
共0篇回复 每页10篇 页次:1/1
我要回复
回复内容
验 证 码
看不清?更换一张
匿名发表 

版权所有 Copyright(C)2013-2020 大连连大生化试剂仪器有限公司

辽ICP备17015610号


点击收缩
返回顶部