Allelopathic Effects of Volatile Monoterpenoids Produced by <Emphasis Type="Italic">Salvia leucophylla</Emphasis>: Inhibition of Cell Proliferation and DNA Synthesis in the Root Apical Meristem of <Emphasis Type="Italic">Brassica campestris</Emphasis> Seedlings

Salvia leucophylla, a shrub observed in coastal south California, produces several volatile monoterpenoids (camphor, 1,8-cineole, -pinene, -pinene, and camphene) that potentially act as allelochemicals. The effects of these were examined using Brassica campestris as the test plant. Camphor, 1,8-cineole, and -pinene inhibited germination of B. campestris seeds at high concentrations, whereas -pinene and camphene did not. Root growth was inhibited by all five monoterpenoids in a dose-dependent manner, but hypocotyl growth was largely unaffected. The monoterpenoids did not alter the sizes of matured cells in either hypocotyls or roots, indicating that cell expansion is relatively insensitive to these compounds. They did not decrease the mitotic index in the shoot apical region, but specifically lowered mitotic index in the root apical meristem. Moreover, morphological and biochemical analyses on the incorporation of 5-bromo-2-deoxyuridine into DNA demonstrated that the monoterpenoids inhibit both cell-nuclear and organelle DNA synthesis in the root apical meristem. These results suggest that the monoterpenoids produced by S. leucophylla could interfere with the growth of other plants in its vicinity through inhibition of cell proliferation in the root apical meristem.     

CpG-ODN对乙肝核酸疫苗及重组疫苗免疫效果的影响

目的研究CpG寡聚脱氧核苷酸(CpGODN)对乙肝核酸疫苗及重组(CHO细胞)疫苗免疫效果的影响。方法核酸疫苗实验组用CpGODN与乙肝核酸疫苗联合免疫BALBc小鼠,阳性对照组注射核酸疫苗,阴性对照组注射质粒pVAX1;重组疫苗分4个实验组,用CpGODN与不同剂量的重组疫苗配伍后分别免疫BALBc小鼠,阳性对照组注射重组疫苗,阴性对照组注射生理盐水。应用ELISA检测小鼠血清中的抗HBs水平。结果CpG与乙肝核酸疫苗组抗体阳转率比阳性对照组提高3倍,抗体水平提高2倍。CpG与重组疫苗组的阳转率和阳转时间均高于或早于CpG与核酸疫苗实验组。加入CpGODN后,即使重组疫苗用量减少3倍,诱导抗体水平和阳转率均无明显改变。结论CpGODN可提高乙肝核酸疫苗和重组疫苗的免疫效果,并使抗体产生的时间提前。     

Properties of cuticular oxidases used for sex pheromone biosynthesis byHeliothis zea

Biosynthesis of the aldehydic sex pheromone components released by females ofHeliothis zea was found to be catalyzed by primary alcohol oxidases residing in the cuticle that covers the glands. Activity, as indicated by conversion of primary alcohol to aldehyde, was as high in cell-free cuticle as it was in intact pheromone glands. Studies indicated that some activity was associated with the surface of the epicuticle and could be removed, into buffer, by sonication. However, the majority of activity lies within the inner epicuticle and exo- and endocuticular layers. The oxidase was not functional in pharate pupae that did not have mature adult cuticle but became functional just prior to adult emergence. The enzyme in individual glands was saturated at alcohol concentrations above 100 n. moles. Nonionic detergents did not affect the activity of the oxidase in the cuticle but treatment with either 7 M urea or 1% SDS resulted in total loss of activity. Studies on the effect of pH indicated an optimum at 6.4; however, activity was high throughout the range of 5–9. The oxidase was functional in both dichloromethane and hexane, suggesting that this enzyme system may have applications for organic synthesis of aldehydes.     

纤维素酶活性测定的生物传感器研究

用TiO2凝胶包埋葡萄糖氧化酶电化学传感器(GOD/TiO2)测定纤维素酶(CE)的活性,即基于电极对纤维素水解后生成葡萄糖的催化氧化。考察了各种实验条件对CE酶活性测定的影响。实验表明,GOD/TiO2电极检测电位为+0.50Vvs.SCE,稳态响应电流与纤维素酶活性在10～300U/L之间有线性关系,该方法可用于微量羧甲基纤维素酶活性测定。     

Access of the substrate to the active site of yeast oxidosqualene cyclase: an inhibition and site-directed mutagenesis approach

A structural model of Saccharomyces cerevisiae oxidosqualene cyclase (SceOSC) suggests that some residues of the conserved sequence Pro-Ala-Glu-Val-Phe-Gly (residues 524-529) belong to a channel constriction that gives access to the active-site cavity. Starting from the SceOSC C457D mutant, which lacks the cysteine residue next to the catalytic Asp456 residue Cys457 has been replaced but Asp456 is still there, we prepared two further mutants where the wild-type residues Ala525 and Glu526 were individually replaced by cysteine. These mutants, especially E526C, were very sensitive to the thiol-reacting agent dodecyl-maleimide. Moreover, both the specific activity and the thermal stability of E526C were severely reduced. A similar decrease of the enzyme functionality was obtained by replacing Glu526 with alanine, while substitution with the conservative residues aspartate or glutamine did not alter catalytic activity. Molecular modeling of the yeast wild-type OSC and mutants on the template structure of human OSC confirms that the channel constriction is an important aspect of the protein structure and suggests a critical structural role for Glu526.     

基于隐马尔科夫模型的DNA序列分类方法

DNA序列分类是生物信息学的一项基础任务,目的是根据结构或功能的相似性预测DNA序列所属的类别。为进行有效分类,如何将序列映射到特征向量空间并最大程度地保留序列中蕴含的碱基间顺序关系是一项困难的任务。为克服现有方法容易导致因DNA序列碱基残缺而影响分类精度等问题,提出一种新的DNA序列特征表示方法。新方法首先为每条序列训练一个隐马尔科夫模型（HMM）,然后将DNA序列投影到由HMM状态转移概率矩阵的特征向量构成的向量空间中。基于这种新的特征表示法,构造了一种 K-NN分类器对DNA序列进行分类。实验结果表明,新型特征表示方法可以较为完整地保留 DNA 序列中不同碱基间的关系,充分反映序列的结构信息,从而有效提高了序列的分类精度。     

基于带约束人工蜂群算法和平均Hausdorff距离的重力匹配方法

重力匹配算法是实现重力辅助惯性导航系统的关键技术之一。但是,传统重力匹配方法存在复杂度高、应用范围小等缺陷,一般很难实现精确、快速匹配的效果。将人工蜂群算法用于重力匹配的搜索过程,并将多普勒测速仪提供的速度信息作为限制条件对蜂群搜索过程进行约束。在此基础上,利用平均Hausdorff距离对匹配结果进行筛选,在重力数据库中重力异常精度一定的条件下,可降低误配率。仿真结果表明,该匹配算法在重力特征显著的区域具有较高的匹配率,可以达到精确、快速的匹配定位,从而实现重力辅助导航。     

猪流感复合多表位核酸疫苗的构建及其表达研究

目的构建猪流感病毒复合多表位核酸疫苗,并研究其表达产物的抗原性。方法以H3、H9亚型流感的HA抗原表位及流感病毒的其它主要抗原(NP、NA、M)表位基因为基础,再附以Kozak序列和适当的酶切位点,设计并合成一段长765bp的复合多表位基因(Epi),其中各表位基本独立,将其克隆入真核表达载体pIRES1neo,构建重组质粒pIRE-Epi。将多表位抗原基因Epi插入到融合表达基因H57中,构建重组质粒pIRE-H57-Epi。将构建的重组质粒在Hela细胞中表达,并用免疫荧光方法初步检测所表达蛋白的抗原性。结果所构建的融合表达载体pIRE-Epi和pIRE-H57-Epi,在Hela细胞中表达出流感病毒多表位抗原蛋白,并具有抗原性。结论已成功构建猪流感病毒多表位基因,有可能成为较理想的猪流感候选疫苗。     

Improvement of cyclodextrin glucanotransferase as an antistaling enzyme by error-prone PCR

In an effort to improve the properties of cyclodextrin glucanotransferase (CGTase) as an antistaling enzyme, error-prone PCR was used to introduce random mutations into a CGTase cloned from alkalophilic Bacillus sp. I-5 (CGTase I-5). A mutant CGTase[3-18] with the three mutations M234T, F259I and V591A was selected by agar plate assay. Sequence alignment of various CGTases indicated that M234 and F259 are located in the vicinity of the catalytic sites of the enzyme and V591 in the starch binding domain E. The cyclization activity of CGTase[3-18] was dramatically decreased by 10-fold, while the hydrolyzing activity was increased by up to 15-fold. These mutations near subsite +1 (M234T) and at subsite +2 (F259I) are likely to alter the enzyme activity in a concerted manner, promoting hydrolysis of substrate while retarding cyclization. The addition of CGTase[3-18] reduced the retrogradation rate of bread by as much as did the commercial antistaling enzyme Novamyl during 7-day storage at 4 degrees C. No cyclodextrin (CD) was detected in bread treated with CGTase[3-18], whereas 21 mg of CD per 10 g of bread was produced in bread treated with wild-type CGTase.     

现代生物工程技术简介

对组成现代生物工程的六大技术系统：基因工程、蛋白质工程、酶工程、细胞工程、微生物工程、生化工程的历史和现状，工作内容及应用前景作了简明的介绍。