Functional Ptoteomic on Antibiotic Resistance and Protective Immunogen of Pseudomonas aeruginosa Outer Membrane Proteins

铜绿假单胞菌是人类的机会致病菌,极少感染健康个体，但对免疫缺陷者，如烧伤、艾滋病、癌症或囊性纤维化等患者则有较高的感染率。铜绿假单胞菌具有内在的耐药性，该菌的耐药机制以及相关疫苗的开发一直以来都备受研究人员的重视。 本文运用蛋白质组学特别是免疫蛋白质组学的方法对铜绿假单胞菌的外膜蛋白进行了研究，对其进行分离和鉴定，在此基础上分析了耐药株外膜蛋白的表达变化以及铜绿假单胞菌外膜免疫原性蛋白。 首先，我们用月桂酰肌氨酸钠的方法提取了铜绿假单胞菌的外膜蛋白，对其进行了2-DE，并通过质谱分析鉴定了其中26个外膜蛋白，构建了铜绿假单胞菌外膜蛋白的分子表达图谱，为下一步的研究奠定了基础。 然后，通过...Pseudomonas aeruginosa, an environmentally versatile bacterium, is an opportunistic human pathogen, infecting healthy individuals rarely, but immunocompromised patients, like burn victims, AIDS-, cancer- or cystic fibrosis-patients at increased risk. This bacterium is high intrinsic resistant to most antibiotics. Thus the understanding of the resistant mechanisms and the development of vaccines ...学位：理学硕士院系专业：生命科学学院生物学系_生物化学与分子生物学学号：20032613

Establishment of Subproteome Map of Pseudomonas aeruginosa OMPs

铜绿假单胞菌是人类重要的病原菌,容易造成免疫缺陷患者如烧伤、艾滋病、癌症或囊性纤维化感染,对该菌外膜蛋白的研究有助于了解该菌的致病机制,为进一步疫苗研制奠定基础.采用月桂酰基氨酸钠的方法提取了铜绿假单胞菌的外膜蛋白,首次建立了外膜亚蛋白质组学表达图谱,鉴定了23个外膜蛋白,占所鉴定蛋白的82.1%.然后对其中的一个外膜蛋白基因PA3988进行了克隆及原核表达与纯化,经免疫小鼠获得抗血清后对外膜蛋白进行双向Westernblotting分析,验证了质谱的结果.这些研究结果对铜绿假单胞菌外膜蛋白的深入研究具有积极作用.Pseudomonas aeruginosa is an opportunistic human pathogen,especially for immunocompromised individuals such as the patients with burn victims,AIDS,cancer or cystic fibrosis.Meanwhile,bacterial outer membrane proteins(OMPs) are important for Gram-negative bacteria because they are involved in adhesion to and invasion of host cells,and in response to the chemical and physical factors of the external milieu.In the current study,the OMPs of P.aeruginosa were investigated with the use of sub-proteomic methodologies by analysis of sodium lauroyl sarcosine-insoluble fraction.Thirty-one proteins were identified,of which 23 were OMPs,accounting for 82.1%.Their function was assigned.Furthermore,PA3988 gene,one of the identified 23 protein genes,was randomly selected for cloning.The cloned gene was expressed for antibody preparing.The prepared antibody was used as the primary antibody in 2-DE Western blotting for confirmation of an identified spot by mass spectrometry.This established complete proteomic profile sheds new insight into the function(s) of E.coli outer envelope.国家自然科学基金973项目(2006CB101807)资

Experimemtal Study on One New Immunostimulated Blue Algae as Additive in Feed for Anguilla japonica Black Fry

免疫增强蓝藻是把胸腺素α1(Tα1)基因转移到蓝藻-聚球藻PCC 7942(Synechococcussp.PCC7942)染色体上后获得稳定表达的转基因蓝藻,经小鼠实验初步表明其有生物学活性并且口服有效.本实验采用该免疫增强蓝藻饲喂日本鳗鲡(Anguilla japonica)黑仔,并采集分离了一种养殖鳗鱼流行性发病弧菌(Vibriosp.),对饲喂了免疫增强蓝藻的黑仔鳗进行感染试验.实验表明饲喂添加免疫增强蓝藻的饲料后,黑仔鳗各检测组织器官Tα1含量显著提高,免疫增强蓝藻提高了试验鱼的免疫能力,增强了抗逆、抗病菌感染能力,并具有一定的促生长作用.免疫增强蓝藻作为鱼类饲料添加剂适口性好,适宜添加量为(0.5~1)×10-6左右.The immunostimulated blue algae is a effectively expressed transgenic blue algae which has translated human thymosin-alpl1(Tα_1) gene to Synechococcus sp.PCC7942.It has been confirmed that immunocompetence is available to mice and can be utilized by oral administration.We fed japonica eel with the immunostimulated Synechococcus sp.as immunostimulated additive,isolated an epidemic pathogenic Vibrio sp.(04~09)from the diseased japonica eel, infected the black fry which were fed with the immunostimulated blue algae with Vibrio sp..The experiment shows that contents of Tα_1 in the tissues and organs of the black fry increased obviously.The immunostimulated Synechococcus sp.can elevate the immune ability of japonica eel,increase the ability of resisting the bacterial pathogens such as Vibrio sp.and environmental stresses,stimulate the growth of japonica eel.The immunostimulated blue algae is an appropriate additive of copulated feed for the rare aquicultured fish and additive dosages is about(0.5~1.0) ×10~(-6)

JUNO Sensitivity on Proton Decay p→νˉK+p\to \bar\nu K^+ Searches

The Jiangmen Underground Neutrino Observatory (JUNO) is a large liquid scintillator detector designed to explore many topics in fundamental physics. In this paper, the potential on searching for proton decay in $p\to \bar\nu K^+$ mode with JUNO is investigated.The kaon and its decay particles feature a clear three-fold coincidence signature that results in a high efficiency for identification. Moreover, the excellent energy resolution of JUNO permits to suppress the sizable background caused by other delayed signals. Based on these advantages, the detection efficiency for the proton decay via $p\to \bar\nu K^+$ is 36.9% with a background level of 0.2 events after 10 years of data taking. The estimated sensitivity based on 200 kton-years exposure is $9.6 \times 10^{33}$ years, competitive with the current best limits on the proton lifetime in this channel

JUNO sensitivity on proton decay p → ν K + searches*

The Jiangmen Underground Neutrino Observatory (JUNO) is a large liquid scintillator detector designed to explore many topics in fundamental physics. In this study, the potential of searching for proton decay in the $p\to \bar{\nu} K^+$ mode with JUNO is investigated. The kaon and its decay particles feature a clear three-fold coincidence signature that results in a high efficiency for identification. Moreover, the excellent energy resolution of JUNO permits suppression of the sizable background caused by other delayed signals. Based on these advantages, the detection efficiency for the proton decay via $p\to \bar{\nu} K^+$ is 36.9% ± 4.9% with a background level of $0.2\pm 0.05({\rm syst})\pm$$0.2({\rm stat})$ events after 10 years of data collection. The estimated sensitivity based on 200 kton-years of exposure is $9.6 \times 10^{33}$ years, which is competitive with the current best limits on the proton lifetime in this channel and complements the use of different detection technologies