固氮菌Klebsiella variicola GN02基因序列及其分泌胞外多糖的相关特性

【背景】一些固氮菌能分泌胞外多糖,与宿主植物的关系密切,在提供氮素和促进植物生长有重要作用。【目的】进行固氮菌Klebsiella variicola GN02的基因序列分析,了解分泌胞外多糖的相关基因和其蛋白的结构特性。【方法】用二代和三代测序技术相结合的方法对GN02菌株进行全基因组分析,并对其分泌的胞外多糖进行理化、结构特性解析。【结果】GN02菌株基因组中含有许多与氮代谢及多糖合成、分泌相关的基因及蛋白,该菌株培养后提取的胞外多糖得率为6.90g/L、分子量1.8×10³Da、比旋度[α]_D²⁵75°、粘度[η]80.28;多糖组成为葡萄糖、半乳糖和甘露糖,为(1→3)及(1→6)糖苷键连接的β构型多糖,具有多糖的红外光谱特征吸收峰。【结论】提供了K.variicola GN02菌株基因组序列,解析了分泌胞外多糖的基因相关蛋白、理化性质及结构特性,有助于进一步了解该固氮菌分泌多糖的机理及为植物促生作用的相关性研究提供基础。     

植物环蛋白的结构

简要介绍了植物环蛋白的定义、结构特点、研究历史、分布、提取分离方法、化学合成与生物合成、生物活性与生物功能.并主要以从紫花蔓地丁(Viola labridorica)中分离得到的六个环蛋白之一,cycloviolacin 02为例介绍通过还原酶解-质谱与二维核磁共振谱结合鉴定环蛋白结构的研究方法.     

Poster Sessions BP02: Oligodendrocytes and Schwann Cells

Neurofibromatosis Type 1 tumors are highly vascularized and contain Schwann cells with hyperactivated Ras. In vitro , the NF1-derived neurofibromin deficient Schwann cells have an angiogenic profile, which favors angiogenesis and sustains the growth of the NF1-derived tumors. This study examined the relationship of the activation state of Ras as it related to the expression of angiogenic and antiangiogenic factors in both cultured NF1-derived Schwann cells and normal human Schwann cells. Western blot analysis of normal human Schwann cells revealed low expression of angiogenic vascular endothelial growth factor (VEGF) as well as low expression of the antiangiogenic pigment epithelium derived factor (PEDF). Relative to normal human Schwann cells, NF1-derived Schwann cells have increased RAS activity and a three-fold increase in VEGF expression. Surprisingly, PEDF was also expressed in the NF1-derived Schwann cells at approximately the same level as VEGF expression. Using a retroviral construct, we introduced the GAP-related domain of neurofibromin into the NF1-derived Schwann cells to reduce the level of activated Ras. Relative to the untreated NF1-derived Schwann cells the Schwann cells expressing the GAP-related domain expressed about one-half the VEGF but twice the PEDF. We conclude that decreasing the Ras activity in NF1-drived Schwann cells will not only decrease proliferation, but also slow tumor angiogenesis due to the decreased expression of angiogenic and increased expression of antiangiogenic factors.     

SP02 particles mediating transduction of a plasmid containing SP02 cohesive ends

SP02 particles that mediate transduction of plasmid pPL1010, a 4.6-megadalton derivative of pUB110 containing an Eco RI endonuclease-generated fragment of SP02 deoxyribonucleic acid that spans the cohesive ends, exhibit three unusual features: the transducing particles have a lower buoyant density than infectious particles; the transduction of pPL1010 occurs at high efficiency; and the transducing activity of the particles is relatively resistant to ultraviolet irradiation when the recipient is recombination proficient. Evidence is presented which indicates that SP02(pPL1010) particles carry the plasmid predominantly as a linear multimer having a molecular mass comparable to that of infectious SP02 deoxyribonucleic acid (ca. 31 megadaltons). The plasmid monomers in the linear multimer appear oriented in the same polarity. The buoyant density difference between infectious and transducing particles appears to be due mainly to the buoyant density difference between pPL1010 (1.699 g/cm3) and SP02 deoxyribonucleic acid (1.702 gm/cm3).     

Naturally Processed Non-canonical HLA-A*02:01 Presented Peptides

Human leukocyte antigen (HLA) class I molecules generally present peptides (p) of 8 to 11 amino acids (aa) in length. Although an increasing number of examples with lengthy (>11 aa) peptides, presented mostly by HLA-B alleles, have been reported. Here we characterize HLA-A*02:01 restricted, in addition to the HLA-B*0702 and HLA-B*4402 restricted, lengthy peptides (>11 aa) arising from the B-cell ligandome. We analyzed a number of 15-mer peptides presented by HLA-A*02:01, and confirmed pHLA-I formation by HLA folding and thermal stability assays. Surprisingly the binding affinity and stability of the 15-mer epitopes in complex with HLA-A*02:01 were comparable with the values observed for canonical length (8 to 11 aa) HLA-A*02:01-restricted peptides. We solved the structures of two 15-mer epitopes in complex with HLA-A*02:01, within which the peptides adopted distinct super-bulged conformations. Moreover, we demonstrate that T-cells can recognize the 15-mer peptides in the context of HLA-A*02:01, indicating that these 15-mer peptides represent immunogenic ligands. Collectively, our data expand our understanding of longer epitopes in the context of HLA-I, highlighting that they are not limited to the HLA-B family, but can bind the ubiquitous HLA-A*02:01 molecule, and play an important role in T-cell immunity.