Homology modeling of anti-parathion antibody and its interaction with organophosphorous pesticides and analogues

The mechanism of specific recognition in pesticide immunochemistry was investigated by computer-based strategy, and a rapid method for the identification of antibody specificity was developed. Based on the previously produced anti-parathion monoclonal antibody (mAb), the DNA sequence was analyzed by polymerase chain reaction (PCR). From the translated amino acid sequences, a three-dimensional structure of the antibody was constructed by homology modeling method, and then it was coordinated by 1 ns molecular dynamics under the explicit solvent condition. The stereochemical property and folding quality were further assessed by Procheck and Profile-3D. The self-compatibility score for the antibody model was 98.7, which was greater than the low score 46.2 and close to the top score 102.6. In addition, parathion and several structural analogues were docked to the constructed antibody structure. The docking results showed that the interaction energy (-40.54 kcal/mol) of antibody-parathion complex was the lowest among all the tested pesticides, which accounted for the high specificity of the antibody to parathion and perfectly matched with the experimental data. Moreover, three residues, Phe165, Asp107 and Thr100 were recognized as the most important residues for antibody reacting with parathion. The interaction energy negatively correlated with the antibody specificity.     

扑草净降解菌降解特性、降解基因保守序列的扩增及同源性分析

从受污染农田中分离到一株补草净降解菌FR，经鉴定属于假单胞菌，考察了温度及pH对其降解能力的影响，确定pH7.0,32℃为最适作用条件；设计引物，通过PCR扩增到一条长度约为500bp的降解基因保守片段，通过对该片段部分序列测定及同源性搜索，证实该片段与atzA,triA等均三嗪化合物水解酶基因具有很高的同源性。     

陕甘宁地区根瘤菌数值分类与DNA同源性分析

在陕甘宁地区根瘤菌资源调查的基础上,选取其中与黄芪、岩黄蓍、木兰、鸡眼草及胡枝子共生的４０株快生根瘤菌和２５株参比菌株,对１３３项表型性状进行聚类分析,在８２％的相似性水平上构成５个不同于已知种的新亚群．对第１、２、３、４亚群ｘ（Ｇ＋Ｃ）／％测定及ＤＮＡ同源性分析结果表明,４个亚群ｘ（Ｇ＋Ｃ）／％含量均处于Ｒｈｉｚｏｂｉｕｍ属范围;各亚群内菌株ＤＮＡ同源性均在７５％以上,每个亚群中心菌株与１３个已知种模式菌株的ＤＮＡ同源性均小于６３％．从而证明,第１、２、３、４亚群为４个独立的快生根瘤菌新种群     

应用紫外及荧光光谱法研究三种醛类污染物对牛腺DNA的损伤作用

应用紫外光谱移动法和溴乙锭荧光法对三种醛类化合物与牛腺ＤＮＡ的加合以及它们引起的ＤＮＡ－ＤＮＡ交联进行了检测。结果表明,甲醛和乙醛能够与ＤＮＡ形成共价加合物,与四种脱氧核苷加合顺序分别为ｄＧ〉ｄＣ〉ｄＡ〉ｄＴ和ｄＡ〉ｄＧ〉ｄＣ〉ｄＴ,丙烯醛与ＤＮＡ的加合不能用紫外光谱移动法测定。甲醛能够显著地引起ＤＮＡ－ＤＮＡ的交联;乙醛也有这种作用,但不显著;丙烯醛的交联作用未测出,可能是由于羰基碳正电性较弱,     

Production of anti-amoxicillin ScFv antibody and simulation studying its molecular recognition mechanism for penicillins

The molecular recognition mechanism of an antibody for its hapten is very interesting. The objective of this research was to study the intermolecular interactions of an anti-amoxicillin antibody with penicillin drugs. The single chain variable fragment (ScFv) antibody was generated from a hybridoma cell strain excreting the monoclonal antibody for amoxicillin. The recombinant ScFv antibody showed similar recognition ability for penicillins to its parental monoclonal antibody: simultaneous recognizing 11 penicillins with cross-reactivities of 18–107%. The three-dimensional structure of the ScFv antibody was simulated by using homology modeling, and its intermolecular interactions with 11 penicillins were studied by using molecular docking. Results showed that three CDRs are involved in antibody recognition; CDR L3 Arg 100, CDR H3 Tyr226, and CDR H3 Arg 228 were the key contact amino acid residues; hydrogen bonding was the main antibody–drug intermolecular force; and the core structure of penicillin drugs was the main antibody binding position. These results could explain the recognition mechanism of anti-amoxicillin antibody for amoxicillin and its analogs. This is the first study reporting the production of ScFv antibody for penicillins and stimulation studying its recognition mechanism.     

家蚕丝心蛋白轻链cDNA和基因调控片段的克隆及序列分析

从家蚕品种菁松×皓月后部丝腺中提取总RNA ,用RT -PCR扩增技术克隆到家蚕丝心蛋白轻链cDNA .序列分析表明 ,该片段全长为 798个碱基 .比较 4种不同品种来源的家蚕丝心蛋白轻链基因外显子区域序列 ,同源性在98.0 %～ 99.4 %之间 ,推测的氨基酸同源性在 98.0 %～ 99.6 %之间 ,4个品种间发生的变异比较一致地集中在不同位置的 8个碱基上 .提取家蚕品种菁松×皓月后部丝腺总DNA ,进行PCR反应 ,获得家蚕丝心蛋白轻链基因调控片段 ,长度约为 1.2kb,测定了其邻近起始密码子一端包括 5 76个碱基的DNA序列 .结果表明 ,该片段包括一些典型的调控元件和多个可能参与丝蛋白基因特异性表达调控的元件 ,该部分序列与J - 139L链基因相应区域同源性高达 98.8% .图 3参 8     

不同浓度甲醛致小鼠肾细胞DNA损伤效应研究

为了探讨甲醛导致生物机体内的DNA损伤效应及其剂量-效应关系,通过不同浓度的液态甲醛对小鼠肾细胞进行染毒,并分别运用了单细胞凝胶电泳实验、荧光检测法实验和KCl-SDS沉淀法实验进行研究.结果显示,甲醛在低浓度(5μmol·L-1)时,具有致DNA断裂的作用;在中等浓度(25μmol·L-1)时,对DNA的损伤作用以DNA-DNA交联为主;在高浓度(125、625μmol·L-1)时,对DNA的损伤作用以DNA-蛋白质交联为主.