玉米ZmPT6基因的克隆及RNAi表达载体的构建

采用PCR方法分别从玉米郑单958基因组DNA及RNA中克隆了菌根共生磷酸盐转运蛋白基因ZmPT6,序列分析表明,ZmPT6基因全长cDNA 1 665 bp,包括一个长97 bp的内含子;编码554个氨基酸,属于Pht1家族成员,由12个疏水的跨膜结构域组成.并构建了35S启动子驱动的植物正义、反义及RNAi表达载体,为下一步玉米的遗传转化及ZmPT6基因的功能研究奠定了基础.     

INO启动子克隆及胚珠特异表达载体的构建

为了观察转基因植株胚珠特异性表达,采用CTAB法提取拟南芥基因组DNA,构建了35S::pCAMBIA1304-L117载体并对番茄进行遗传转化。根据已报道的INO启动子序列设计并合成一对特异引物,通过PCR对启动子进行扩增、连接并对其进行测序,结果表明：与GenBank中已注册的INO启动子序列的同源性为100%,并且构建了以INO代替CAMV35S启动子的pCAMBIA1304-L117植物表达载体,采用冷冻法将其转入根癌农杆菌EHA105。成功构建了35S::pCAMBIA1304-L117载体,并为进一步检测奠定了基础。     

黄瓜芽黄突变相关基因ClpP克隆及植物表达载体构建

为了获得黄瓜中ClpP基因的cDNA全序列,研究其与黄瓜芽黄突变现象的关系,本研究采用TRNzol法提取芽黄突变的黄瓜叶片总RNA,并以其为模板反转录合成cDNA第一链。根据NCBI预测的黄瓜ClpP基因序列设计并合成1对特异引物,通过PCR扩增得到目的条带后测序,并构建植物表达载体;成功获得黄瓜中ClpP基因的cDNA全序列,并提交到NCBI。该基因编码区全长495 bp,共编码氨基酸158个。预测其理论等电点为5.22,理论蛋白质分子量为41.00356 KDa,编码的蛋白包含S14_ClpP_2保守结构域,无信号肽。通过与其他植物Clp蛋白酶的氨基酸序列比对发现与毛茛属植物的Clp蛋白酶同源性较高。并成功构建了以CaMV35S为启动子的植物表达载体pBI121-ClpP。黄瓜中ClpP基因的cDNA全序列的获得说明该基因在黄瓜中确实存在,这是首次克隆得到了黄瓜中的ClpP基因的cDNA全序列。     

柚CCoAOMT基因的克隆与超表达载体构建

为探明咖啡酰辅酶A-O-甲基转移酶（CCoAOMT）在柚[Citrus maxima (Burm.) Merr.]果实发育中对木质素单体合成的影响,克隆了柚CCoAOMT基因ORF序列并构建超表达载体转化至农杆菌。采用Trizol法提取柚汁胞总RNA,经反转录得到cDNA,根据已知柚CCoAOMT片段序列,设计其ORF引物。PCR扩增得到柚CCoAOMT基因ORF序列,长度为744 bp,编码247个氨基酸,与龙眼(Dimocarpus longan)、麻疯树(Jatropha curcas)、杨树(Populus balsamifera)的CCoAOMT基因同源性大于90%。经软件预测,该酶定位于质膜的可能性最大,不具有跨膜结构。将柚CCoAOMT基因正向导入植物过表达载体p1301,位置在CaMV35S启动子的之后,并将载体转化至农杆菌EHA105。本研究得到的基因片段包含柚CCoAOMT基因ORF,成功构建植物超表达载体p1301-cmCCoAOMT,并导入农杆菌。含p1301-cm CCoAOMT的农杆菌株可转化多种植物受体,为深入研究该基因功能奠定了基础。     

番茄转录因子LeMADS-MC正反义植物表达载体的构建

为进一步阐明转录因子家族MADS-Box对番茄果实成熟的调控途径,本研究利用聚合酶链式反应技术(PCR)从番茄(Solanum lycopersicum)cDNA中克隆LeMADS-MC基因和LeMADS-antiMC基因核心片段,将这2个基因通过酶切分别正向连接到pCAMBIA1300-221载体和反向连接到pCXSN载体中,并用冻融法将重组载体导入农杆菌中。获得的重组载体分别通过PCR及酶切鉴定符合预期结果且测序结果与NCBI同源性高达100%。结果成功构建了适合于番茄农杆菌遗传转化的植物表达载体,为下一步通过LeMADS-MC基因研究果实成熟衰老机理奠定了物质基础。     

马铃薯晚疫病菌RxLR效应因子RD24基因克隆及其PVX表达载体构建与鉴定

马铃薯晚疫病病原菌致病疫霉菌(Phytophthora infestans)在侵入寄主细胞的同时分泌效应因子(Effector)，该类蛋白在晚疫病致病过程中发挥着关键作用，是抑制植物免疫的重要因素。PITG_RD24是马铃薯晚疫病菌位于细胞核核的一个重要效应因子。为了快速鉴定效应因子RD24在马铃薯中瞬时表达的状况及抗性情况，构建效应子RD24的PVX表达载体是十分必要的。本研究利用PCR、酶切、分子克隆等生物学技术，将RD24片段连接插入至PVX（pGR106）载体中，然后用热击法将连接产物转化至大肠杆菌DH10B感受态细胞中。通过PCR筛选、质粒提取、测序比对，结果表明成功获得含有目的表达载体pGR106·RD24的质粒，可为下一步筛选马铃薯广谱抗病基因研究提供必备载体。     

江汉平原土壤有机碳含量高光谱预测模型优选

研究探讨了贫瘠地区低有机碳含量条件下,不同光谱预处理与建模方法用于土壤有机碳估测的最佳组合,对贫瘠土壤属性信息快速获取和精确农业发展具有重要意义。以江汉平原不同利用条件下的土壤为研究对象,使用可见光/近红外高光谱技术,结合包括Savitzky-Golay平滑(SG)、一阶导数(FD)、多元散射校正(MSC)在内的光谱预处理方法,分别建立用于估测土壤有机碳(SOC)含量的多元线性回归(MLR)、主成分回归(PCR)、偏最小二乘回归(PLSR)和支持向量机回归(SVMR)模型。结果表明：不同建模方法预测精度差异明显,PLSR和SVMR的预测结果优于MLR和PCR;不同预处理方法对模型的预测精度亦有较大影响,表现为MSC＞FD＞SG;基于FD和MSC组合预处理的SVMR模型的预测能力最好,R2=0.84,RPD=2.50,满足土壤有机碳的预测。有机质含量大于2%并不是建立优质模型的必要条件。     

EXISTENCE OF A BASIC OPTIMAL SOLUTION FOR LINEAR VECTOR EXTREME VALUE PROBBEM

In this paper, the concept of basic for linear vector extreme value problem is introduced in the partially ordered vector space. Some properties and results of the linear programming are generalized. And the existaence therom of basic optimal solution is proved.     

A Vector Quantification Image Coding Scheme Based on Wavelet Transform

The paper proposes an image compression and coding scheme based on discrete wavelet transform (DWT), which makes the best of the relativities among original image and coefficients of wavelet, the relativities among pixels of subimages, the relativities among directions of subimages. Human visual specialities were considered, so the compression scheme leads up to make full use of the statistical redundancy and visual redundancy of image. To get high compression ratio and good quality of image, the coefficients of image working on the quality of image greatly were reserved accurately, but the coefficients of image working on the quality of image slightly were quantified roughly. Also, the regrouping and coding of the coefficients could be used universally. The experimental results show that the compression method is provided with simply calculating, little time for coding and decoding and satisfying quality companied with high compression ratio.     

Design of Bootloader for Embedded Systerm on S3C4510B

Recent years, there emerged many new types of 32-bit embedded system services. But due to the non-commonality of 32-bit embedded system, the designers of every system service should design particular boot software. The authors analyze the characteristic of 32-bit microprocessor S3C4510B , and the basic structure of embedded system and the characteristic of boot. The authors discuss the realization process of remap during the boot process and the management of abnormal vector in boot code. At last, they design boot program based on S3C4510B embedded system with good modularity and transplantablity. The test result shows good stability and effectivity of this design.     

ZmPti1基因正义和RNAi表达载体的构建及其转化玉米的研究

构建了ZmPti1基因的正义表达载体和RNAi载体,以bar基因为抗性筛选标记,通过花粉管通道法将构建的两个表达载体分别转化到玉米自交系178.收获的种子播种于营养钵中,出苗后,经0.1%的草丁膦筛选得到40株草丁膦抗性植株,进一步用PCR鉴定与PCR-Southern检测得到33株转基因植株,其中15株是转化正义表达载体的转基因植株,18株是转化RNAi表达载体的转基因植株.并对转化方法和ZmPti1基因的功能进行了讨论.     

语谱特征的身份认证向量识别方法

针对采用梅尔频率倒谱系数（mel-frequency cepstrum coefficient，MFCC）作为身份认证向量（identity vector，i-vector）进行说话人识别存在语音信息不全的问题，提出一种基于语谱特征的身份认证向量识别说话人的方法。语音信号经过预加重、分帧加窗预处理之后，通过短时傅立叶变换转换成语谱图，语谱图被提交到高斯通用背景模型，在高维均值超向量空间中选择合适的低维线性子空间流型结构以构造符合正态分布的向量作为身份认证向量。这些获取的身份认证向量经过线性判别性分析实现降维并存储。最后采用对数似然比（log-likelihood ratio，LLR）方法对训练和测试阶段的i-vector进行评分，完成说话人识别。以TIMIT数据库为标准的数值实验结果表明，相比采用MFCC作为特征的识别方法，研究的等错误率（equal error rate，EER）更低。     

Geometric Parameters Analysis of the Punch in Stamping Die of Thin wall Pipe with Arc shaped End

The punching technology of thin wall pipe with arc shaped end is described.The geometric parameters of the punch are also analyzed by vector analysis method.By minimizing the deformative force,the formula on how to calculate the parameters is derived.The result is proved to be correct by actual experiment.     

Simulation Research of Control Parameters in Induction Motor Vector Control System

In order to obtain the effect rules of control parameters of speed regulator on performance indexes of induction vector-controlled system and the setting rules of control parameters of speed regulator,according to rotor flux oriented control principle,a vector control model of induction motor is established and simulated with the software of Matlab/Simulink.This vector control system is proved that it possesses good static and dynamic performance.This paper analyzes the effect of control parameters' variations of speed regulator on performance indexes and gives simulation results.     

TMV、CMV双价抗性RNA沉默表达载体的构建

本研究以在植物体内转录出RNA沉默的高效诱导因子双链RNA（dsRNA）为目标,根据已报道的TMV ΔMP和CMV ΔRep的核苷酸序列设计特异性引物,分别从质粒pBIN-TMV ΔMP(i/r)、pBIN-CMV ΔRep(i/r)扩增MP基因、Rep基因,并在pGEM-T Easy 载体上将2片段串联起来,再用PCR扩增串联好的正反向MP-Rep基因。正向MP-Rep基因用BamHI和Kpn I切下,将所得基因片段与用同样酶切开的含内含子的pKAN-In相连,取名+pKAN;用同样的方法,将反向MP-Rep基因用Xba I切下,将所得基因片段与用同样酶切开的+pKAN相连,取名pKAN-In-MP-Rep,再用Not I将包括Intron和正反向MP-Rep基因在内的片段切下,将其定向插入同样酶切开的pART27上,获得重组质粒pART27-In-MP-Rep。获得的载体可以应用于植物转基因抗病毒育种工程中。     

Product knowledge documents retrieval based on hybrid semantic model

An approach based on hybrid semantic model (HSM) was proposed to the solve problem raised in the retrieval process of product knowledge documentation. It expands the traditional user query to a semantic set composed of user preference, context and query, while representing the knowledge documents and user interest with an ontology based fuzzy concept. The leaves in the ontology are selected as components of the document concept vector with the weight determined by the depth of the concept in the ontology graph, the quantity of the information contained, and occurrence in the document and the whole repository. Furthermore, ontology is used to express context and query, and to construct a user preference model. Different relevancy computation methods are adopted for different retrieval models. The semantic similarity between query or user preference and documentation is computed by cosine method. The semantic similarity of context is estimated by the concept distance in the concept hierarchy. Finally, the method is shown by experimentation to be more effective than the classic vector space method.     

植原体病害的传播、流行和防治研究进展

植原体是一类通过叶蝉、飞虱、木虱和蝽象等韧皮部取食的半翅目昆虫传播的植物病原细菌,能够引起世界范围内1000 多种植物病害。因其经济上的重要性,植原体病害被广泛而深入的研究。本研究归纳了植原体病害的传播和流行研究的最新进展,总结了植原体病害防治的方法和策略,分析了植原体病害的传播过程及其影响因素、植原体—寄主植物—介体昆虫的分子互作、植原体病害流行的要素和影响因素。此外,还指出了当前研究存在的问题,展望了今后植原体病害研究的重点领域。     

Cloning of aspergillus niger F246 phytase gene and its a new type of expression vector construction

This research amplified the phytase gene of Aspergillus niger F246 by the polymerase chain reaction(PCR), which is selected and identified by ourselves.Then the product of PCR was inserted respectively into the lactobacillus expression vector pMG36e to construct the new type of expression vector pMG36e-phyA. The genomine DNA of the Aspergillus niger F246 strain is used as the template for PCR,and the PCR product was subcloned into pMD18T. After sequncing the coding region and analying the sequnce of amino acids, the product of PCR was cutted from pMD18T and subcloned into pMG36e for the engineering strain of Lactobacillus MG1363-pMG36e-phyA. The construction of new recombinant expression vector pMG36e-phyA for the phyA gene lays the base of the study on obtaining large and high active phytase and developing the new microbial ecologicalagent.     

DREB基因双T-DNA植物表达载体的构建及验证

W34基因经pGEM-T-EASY载体,克隆到pBDREB载体EcoRI位点,得到在Amp平板上生长的转化子pBW34-DREB质粒;质粒pBW34-DREB、pSPROK、pBlueks分别经Pst I／KpnI、KpnI／EcoRI、PstI／EcoRI双酶切,三片段连接构建成中间表达载体pBWD-Tnos;再用SmaI／EcoRV双酶切pBWD-Tnos。将回收的DREB基因的完整的表达结构元插入到经SmaI酶切的质粒载体pCDMAR-Hyg中,构建成双T-DNA植物表达载体pCDMAR-pWDT-Hyg,大小为13．59kb,经酶切鉴定,证明构建的载体与预期设计的一致。将此双T载体用农杆菌介导法转化玉米胚性愈伤组织,期望得到无选择标记的安全的转基因植物。