Preparation and Application of Soluble Human Squamous Cell Carcinoma Antigen Expressed by Escherichia coli

旨在建立基于大肠杆菌表达系统的高效可溶性表达人鳞状上皮细胞癌抗原(SCCAg)方法,获得具有较好活性的重组SCCAg抗原并应用于建立抗原检测方法; 。基于pGEX-6P-l载体和大肠杆菌E. coli; ER2566菌株开展重组SCCAg抗原可溶性表达纯化方法研究,评价纯化抗原活性,筛选特异性单克隆抗体,初步建立并评价SCCAg抗原检测方法。结果; 显示,pGEX-6P-l载体和E coli; ER2566菌株可用于建立较高效的可溶性表达和纯化SCCAg抗原的方法,获得了具有较高纯度和活性的重组SCCAg抗原,筛选获得特异性单克隆抗体并; 初步建立了 SCCAg管式化学发光检测方法。建立了有效的基于大肠杆菌表达系统的可溶性表达和纯化SCCAg的方法。The aims of this study are to establish a method for efficient soluble; expression of human squamous cell carcinoma antigen(SCCAg ) based on; Escherichia coli expression system and obtain the recombinant SCCAg; antigen in fine activity, then apply it in the detection method; establishment of antigen. The study on the method of soluble expression; and purification of recombinant SCCAg antigen was conducted based on; pGEX-6P-l vector and E. coli ER2566 strain. The activity of the purified; antigen was evaluated by Abbott Kit and the specific monoclonal antibody; was screened by indirect ELISA. It was proved that PGEX-6P-1 vector and; E. coli strain ER2566 could be used to establish efficient soluble; expression and purification method for recombinant SCCAg antigen.; Moreover, the recombinant SCCAg antigen was proved to be in high purity; and activity. Thus,the SCCAg detection method of chemical luminous tube; was established with the specific monoclonal antibodies. In conclusion,; an effective method for the expression and purification of SCCAg, which; is based on the E. coli expression system, is established.国家高技术研究发展计划(863计划

Cloning and Characterization of Na~+/H~+ Antiporter Gene (nhaA) from Pseudomonas sp.cn4902

根据3种生物的Na+/H+逆向转运蛋白基因(nhaA)的两端序列设计引物,利用PCR从假单胞菌 (Pseudomonassp.cn4902)中克隆得到一结构基因。该基因长1089bp,编码362个氨基酸,与E.coliK12的 nhaA基因的同源性高达97.0%。将该结构基因与pBV220构建成重组载体pBVA。SDS PAGE电泳表明:含 pBVA的转化子产生较高浓度的分子量约为41kD的蛋白,与预期相符。在含NaCl1.0mol/L的培养基中生长达 到平衡期时,转化子的菌浓度约是对照的2.3倍。经原子吸收光谱测定,转化子细胞质中Na+浓度仅为对照菌的 60.4%。SDS PAGE电泳表明该基因的表达蛋白位于细胞膜(壁)上。提纯外源基因表达蛋白并对其N端8个氨 基酸进行测序,与nhaA基因推测的氨基酸序列完全相符。这些实验证实,克隆得到的基因是假单胞菌的nhaA基 因。该基因已经在GenBank登记,收录号为AY643494。 【英文摘要】 According to the sequences of the gene nhaA coding for Na~+/H~+ antiporter,a structural gene was cloned from Pseudomonas sp.cn4902 by PCR reaction with a set of primers.It was 1 089 bp in length and codes for 362 amino acids sharing homology with the gene nhaA of E.coli K12 as high as 97.0%.It was inserted into plasmid pBV220 to form a high level expression reconstruction plasmid pBVA.So an overexpression 41 kD protein band could be found in the lane of transformant harbored with pBVA after SDS-PAGE electro...福建省科技计划重点资助项目(编号:2003N053);; 厦门大学生命科学学院细胞生物学与肿瘤细胞工程教育部重点实验室项目(编号:2004106)~

HPLC and MALDI-Tof-MS Analysis of Hydrolysates of Xylan

利用HPlC结合MAldI-TOf-MS对1-苯基-3-甲基-5-吡唑啉酮(PMP)衍生化后山毛榉木聚糖水解产物进行分析,检测到了难以获得标准品对照的木聚糖水解产物。结果表明,稀硫酸水解山毛榉木聚糖的主要水解产物有木糖和4-O-甲基葡萄糖醛酸-木糖(b2),以及少量4-O-甲基葡萄糖醛酸(b1)。内切重组木聚糖酶An Xyn10C水解山毛榉木聚糖产生木糖、木二糖和4-O-甲基葡萄糖醛酸-木三糖(b3),而内切重组木聚糖酶HO Xyn11A水解山毛榉木聚糖主要产生木糖、木二糖、木三糖、4-O-甲基葡萄糖醛酸-木四糖(b4)和4-O-甲基葡萄糖醛酸-木五糖(b5)。基于PMP柱前衍生化的HPlC结合MAldI-TOf-MS方法能高效地分析复杂的木聚糖水解产物。The hydrolysis end products of beechwood xylan,which were released by sulfuric acid or enzymes and then labeled at their reducing ends with 1-phenyl-3-methyl-5-pyrazolone( PMP) derivatization,were analyzed by HPLC assisted with MALDITof-MS.Some of the hydrolysates,which were lack of related commercial available standard substances,were determined.It was found that the xylose and 4-O-methyl-glucuronic acid-xylose( B2) were the main products with minor amounts of 4-Omethylglucuronic acid( B1) in the hydrolysates of beechwood xylan by sulfuric acid.Recombinant endo-β-1,4-xylanase An Xyn10 C released xylose,xylobiose,and 4-O-methyl-glucuronic acid-xylotriose( B3) as the main hydrolysates from beechwood xylan,whereas recombinant endo-β-1,4-xylanase Ho Xyn11 A released xylose,xylobiose,xylotriose,4-O-methyl-glucuronic acidxylotetrose( B4) and-xylopentaose( B5),and aldohexaouronic acid.These results revealed that HPLC assisted with MALDITof-MS based on PMP derivatization was a very useful and robust method for the determination of products in hydrolysis of xylan.国家自然科学基金资助项目(31170067); 厦门市海洋经济发展专项资金项目(14GZP59HJ29); 福建省海洋高新产业发展专项项目(闽海洋高新[2014]25号

Analysis of the nutrient distribution features and affecting factors in the Jiulongjiang estuary

根据2009、2010年“丰水期“和“枯水期“四航次九龙江河口混合区的调查资料,且结合历史资料对营养盐含量及分布特征、周日变化特征进行了统计和相关分析,研究了九龙江流域营养盐输入海洋的变化过程,探讨九龙江河口营养盐伴随潮汐变化,以及河口混合过程中的生物地球化学行为。调查期间溶解无机氮、硅和磷含量的平面分布呈现出由径流冲淡水高值向河口外海端递减的变化趋势;在涨潮时,河口区感潮段高溶解无机氮、硅、磷营养盐的陆源冲淡水与低溶解无机氮、硅、磷营养盐外海水相遇,随着外海水的侵入,外海水的作用逐渐加强,在稀释混合过程中呈现出无机营养盐逐步降低的变化趋势,退潮时则相反;营养盐在这复杂的河口过程中往往表现出在水动力的作用下稀释混合是主要过程,无机氮和活性硅酸盐在河口稀释混合过程中呈现保守性特征,活性磷酸盐在河口转移(补充)过程的行为复杂化,呈现缓冲作用为主。Based on the historical data and the data from four surveys in the wet season and dry season of 2009 and 2010 in the Jiulongjiang estuary,the concentration and distribution of nutrient were discussed in the mixing process.Furthermore,the daily variation of nutrient concentration with the tide was also discussed.The results showed that:(1) the concentration of total dissolved inorganic nitrogen,silicate and phosphate all decreased gradually from the river runoff to seawater;(2) in high tide,in the mixing process of the runoff with high nutrient concentration and the seawater with low concentration of nutrient,the concentration of nutrient decreased due to seawater.On the contrary,during the ebb,the concentration of nutrient increased due to the runoff;(3) in the mixing process in Jiulongjiang estuary,nitrogen and silicate were both conservatively diluted,but phosphate was buffered because of the complicated transfer and complement of phosphate.国家海洋局公益性项目(200805064);国家海洋局第三海洋研究所基本科研业务费项目(海三科2009021);海三科200900

ST段抬高型急性心肌梗死院前溶栓治疗中国专家共识

急性心肌梗死仍然严重威胁我国人民健康,在我国广大城乡地区,形势更为严峻[1,2]。及时救治急性心肌梗死患者,降低死亡率和保护心脏功能刻不容缓。鉴于我国的实际情况,院前溶栓治疗在大城市以外的城乡地区具有重要意义。为此,中国医师协会胸痛专业委员会及中国医学救援协会心血管急救分会专门组织有关专家制订了本共识,旨在帮助院前医疗急救人员对急性心肌梗死患者选择最佳

Identification and preliminary analysis of a novel full-length cDNA encoding retinoid X receptor 2 from Schistosoma japonicum

通信作者 E-mail: [email protected][中文文摘]目的克隆编码日本血吸虫视黄酸X受体2(SjRXR2)蛋白的全长cDNA,并对其进行初步研究。方法利用cDNA末端快速扩增技术(RACE)获得SjRXR2蛋白全长编码cDNA。利用生物信息学技术,对基因结构进行初步分析。利用实时荧光定量(Real time)PCR技术对该基因在日本血吸虫不同时期虫体中的转录情况进行分析。应用在线抗体表位预测软件获得SjRXR2配体结合区抗原性较强的一个多肽序列,合成该多肽片段,并免疫小鼠制备抗血清。利用Western blot技术分析该蛋白在日本血吸虫中的表达。结果采用RACE技术成功获得了SjRXR2蛋白全长编码cDNA,总长度为5 960bp,其完整开放阅读框为4 308 bp,编码1 435个氨基酸,预测分子量为159 kDa。生物信息学分析表明该基因编码的蛋白质序列具有核受体家族2的典型结构域特征,且与曼氏血吸虫RXR2有较高的相似性。Real time PCR分析表明,该基因在21、42 d龄日本血吸虫虫体内有较高的转录水平。Western blot分析表明,小鼠SjRXR2多肽免疫血清可特异性识别日本血吸虫虫体150 kDa蛋白。结论成功获得了编码SjRXR2蛋白的全长 cDNA，并制备了针对该蛋白的特异性多克隆抗体，为进一步研究该蛋白的功能奠定了基础。 .国家自然科学基金（31172315）；公益性行业（农业）科研专项（20090303036

城乡链接与农民合作

2009-2010 > Academic research: refereed > Publication in refereed journalVersion of RecordPublishe

1950-2007年黄河入海水沙通量变化趋势及突变特征

基于1950-2007年黄河利津站水沙数据,采用Mann-Kendall检验以及Mann-Whitney-Pettitt(MWP)与贝氏变点分析方法来分析黄河入海水沙通量变化规律,结果表明:黄河全年入海径流通量与泥沙通量分别以-8.1139亿m~3/a和-0.2285亿t/a速率显著减少,汛期变化幅度大于非汛期,尤以泥沙通量为甚;全年以及汛期和非汛期入海径流通量与泥沙通量时序均存在显著转折,且各自变点出现时间不完全一致,全年入海径流通量与泥沙通量时序转折分别发生于1968年、1985年、2002年和1968年、1985年、1996年;入海水沙通量变化趋势与时序变点与流域自然因素变化与人类活动影响密切相关,部分变点出现时间与人类活动介入相吻合