法氏囊活性肽在大肠杆菌中的克隆与表达

法氏囊活性肽（Bursin,BS)作为一种免疫增强剂，临床应用广泛，为了大量制备BS，按大肠杆菌惯用密码子合成BS基因。并用色氨酸将5个BS基因串联，克隆于质粒pU57的SmaI位点，经测序证明序列正确后，以PCR扩增，克隆于表达载体pBV220的EcoRI,HinDⅢ位点，温敏诱导表达，其表达水平占全菌总蛋白的13.4%。     

实广义正定矩阵的若干性质

矩阵∈AR~(n×n)称为实广义正定矩阵,如果对任意非零向量 X∈R~n,有XTAX>0成立。本文讨论了矩阵的kronecker积。Hadamard积和矩阵乘积的正定性,给出相应一些性质。     

Kinetics of NDV specific antibodies in chickens—V. Analysis of frequency distributions of antibody titers against Newcastle disease virus by investigation of random samples in chicken flocks

Chicks and chickens maintained under commercial conditions were vaccinated against Newcastle Disease via drinking water. Prior and after different times of vaccination blood samples were drawn from different numbers of birds and checked for HI antibodies. The modes of distribution of the antibody titers within the random sample were assayed. The following results were obtained:

1. 1. On the basis of the distribution of the serum titers can be concluded whether the antibody level within a flock is increasing or decreasing.

◦ —A right steep asymmetry can be observed up to 20 days post vaccination.

◦ —In the phase of maximal antibody levels an almost symmetric distribution of the titers is present.

◦ —In later times (more than three weeks p. vacc.) the distribution shows a left steep asymmetry (Poisson distribution).

2. 2. A poisson distribution is also observable during the elimination of maternal antibodies of chicks until complete elimination.

3. 3. The mode of distribution of the HI titers in sera of day-old chicks correlates with the mode of distribution of the dams. Therefore, conclusions are possible from the status of the chicks to the dams and reverse.

4. 4. Factors which interfere with the mode of distribution are:

◦ —Two or more peaks after vaccination of chickens. This indicates an uneven immune response within the flock.

◦ —Distributions with several peaks may also occur if flocks are composed of day-old chicks from parent flocks with different levels of antibody titers.

The value of vegetation cover for ecosystem services in the suburban context

Latin-American cities can be characterized by dynamic processes of urbanization that encroach upon the natural and semi-natural surrounding landscapes. Our study presents the effects of landscape development, transformed from semi-natural conditions into a mostly disperse suburban settlement. We explore the impact that this transformation has had on this context by three ecosystem services that regulate rainwater runoff, enhance microclimate conditions and help to improve air quality by monitoring vegetation cover. We have designed a spatio-temporal hierarchical analysis which employs remote sensing techniques to capture the structural changes of this landscape over long, medium and short term scales on two spatial levels. This methodological approach was tested in the Metropolitan Area of Santiago (MAS) as case study area. Despite of the increase in impervious surfaces due to urban processes, there has also been an increase in vegetation cover, which has led to an improvement in the provision of the above-mentioned ecosystem services. Hence, if diverse urbanization processes continue and they are coupled with an increase in vegetation cover, the provision of ecosystem services could also expand. This phenomenon can be observed in some areas, where public and private green spaces are created and maintained. Our data analyses give evidence that certain types of suburban areas which increase the share of vegetation cover can provide daily ecological benefits for urban neighborhoods, and beyond, for adjacent areas. Moreover, suburban development can successfully provide ecological benefits to citizens. Such processes can only be ecologically sustainable if the composition of vegetation is well-adapted to the regional climatic conditions.     

苦瓜转录因子McEIL2的克隆及其表达分析

本研究从已构建的苦瓜果实均一化文库筛选得到一个EIN3-like同源EST序列,结合RACE技术,克隆得到EIN3基因c DNA全长序列,命名为Mc EIL2(Gen Bank登录号:KF595122)。该基因全长2 122 bp,开放阅读框1 890 bp,编码629个氨基酸,预测该蛋白的分子量为71.58 k D,与黄瓜Cus EIN3、甜瓜Cm EIL1、苜蓿Mt EIL1、绿豆Vr EIL1和番茄Le EIL1的蛋白同源性分别为89.61%、78.37%、71.23%、69.09%和65.66%。Mc EIL2基因的编码蛋白亚细胞定位于细胞核,荧光定量分析表明Mc EIL2基因表达量在幼果期先强后弱,绿熟期最低,破色期表达量大幅增加至最高随后下降,推测该基因参与苦瓜果实成熟软化调控过程。本研究初步明确了Mc EIL2基因在苦瓜成熟过程表达模式,可为今后进一步揭示苦瓜果实成熟软化的分子机制奠定基础。     

普通小麦春化基因VRN1 RNA干扰载体构建及转基因小麦获得

为进一步探究春化基因VRN1在小麦发育进程中的功能,利用荧光定量PCR分析了VRN1基因在不同发育特性小麦品种新春2号、京841中的表达情况。结果表明,随着生育进程的推进,VRN1基因在新春2号叶片和茎尖中表达量均呈上升趋势,VRN1基因在京841叶片中呈波动上升趋势,在茎尖中表达量趋于0;以p FGC5941载体为基础构建含有VRN1反向重复序列的RNA干扰载体,利用农杆菌介导的茎尖转化法转化新春2号获得了再生植株,并通过PCR法检测获得了转基因阳性植株,为从分子水平上实现小麦发育特性遗传改良和创育小麦新种质奠定了基础。     

高粱光敏色素A基因的克隆、蛋白结构与长短日照诱导表达模式

光敏色素是一类红光/远红光受体,对植物光形态建成、外部形态和生理功能等方面起着重要的调节作用。为明晰光敏色素A在调控高粱光形态建成发育过程中的作用,本研究比较分析光温敏感程度不同的高粱品种Btx623、Rio、Tx430和LH645的PhyA基因序列和PHYA蛋白的保守结构域,利用qRT-PCR明确长短日照条件处理下PhyA基因的表达模式。结果表明,Btx623、Rio和Tx430之间PhyA基因序列存在非同义突变,LH645在PhyA基因的第7个外显子上插入了5 bp碱基序列;高粱PhyA蛋白中含有1个PAS-2结构域、1个GAF结构域、1个Phytochrome结构域、2个PAS结构域、1个组氨酸激酶A结构域和1个类似组氨酸激酶的ATP激酶结构;遗传进化树分析表明,单子叶和双子叶植物PhyA蛋白明显聚为2大类,高粱PhyA、谷子PhyA和玉米PhyA1、PhyA2聚为一个亚类,相互间亲缘关系较近,与水稻PhyA同源性相对较远;在短日照(SD)条件下各时期PhyA基因转录水平变化显著,Rio PhyA基因转录水平较Btx623和Tx430变化明显,LH645 PhyA基因转录变化不显著,在长日照(LD)条件下Rio PhyA基因转录水平低于Btx623和Tx430,LH645 PhyA基因转录水平变化不显著。以上结果表明,PhyA受光周期诱导,推测其在高粱光形态建成中起重要调控作用。本研究为进一步解析PhyA基因功能及其在调控高粱光形态建成发育过程中的作用提供理论基础。     

砂藓(Racomitrium canescens)抗旱相关基因RcAOS2的克隆及表达分析

丙二烯氧化物合酶(allene oxide synthase, AOS)是茉莉酸合成途径中关键酶,在茉莉酸合成中具有重要的作用。本研究在砂藓(Racomitrium canescens)干旱胁迫转录组数据中发现一个干旱胁迫下表达量上调的AOS基因,将其命名为RcAOS2,我们对RcAOS2的开放阅读框进行了克隆及相关的生物信息学预测分析,并对RcAOS2在砂藓快速脱水和复水过程中的表达量变化进行了检测。结果显示,该基因cDNA全长1 474 bp,开放阅读框长516 bp。RcAOS2编码蛋白质的相对分子质量为4.36 kD,等电点为5.11,不稳定系数为55.02,为不稳定蛋白质;脂肪系数为19.19;RcAOS2蛋白质疏水性较强,属于疏水性蛋白质,存在信号肽,不属于跨膜蛋白质。系统进化分析表明,RcAOS2蛋白质与其他物种AOS蛋白质亲缘关系较远,是具有AOS序列特征的新类型。实时荧光定量PCR分析结果表明RcAOS2基因在砂藓干旱和复水过程中呈现差异性表达,推测RcAOS2可能参与砂藓的干旱胁迫响应和复水回复过程。本研究成果为进一步研究砂藓抗旱基因功能提供理论基础和备选基因资源。     

Serum amyloid A (SAA) mRNA expression in chicken and quails in response to bacterial stress

Monitoring of acute phase proteins such as serum amyloid A at gene expression level may provide quick information about immune status of the host and its susceptibility towards common infections. Present study was carried out to evaluate and compare the mRNA expression of SAA gene in Rhode Island Red chicken (RIR) and Japanese quails using real time PCR analysis in response to inactivated Salmonella gallinarum culture. The results showed that expression of SAA gene was approximately 17–33 folds higher in case of birds administered with bacterial culture when compared to un-inoculated controls and expression was higher and quicker in case of quails than RIR chicken. The SAA genes from chicken and quail were cloned and upon sequence analysis it was observed that deduced amino acid sequence of SAA from chicken and quails were having approximately seven percent variation which might have significance in function of this protein in these species.