用mRNA差异显示技术分离盐胁迫下小麦耐盐相关cDNA

目的用mRNA差异显示技术分离小麦盐胁迫应答cDNA,为进一步研究小麦耐盐机理奠定基础。方法将小麦(Triticum aesticum L.)耐盐品系宝丰7228r种子分别置于含NaC l 0‰和12‰的Hoagland培养液中生长,7d后分别取叶片,提取总RNA并分离出其中的mRNA,通过锚锭引物O ligo dT10GC反转录和9个10核苷酸随机引物进行PCR扩增。结果DDRT-PCR结果显示,有4个差异DNA片段只在盐胁迫的小麦耐盐品系基因组中表达,而在对照中没有出现。这4个差异cDNA片段分别命名为ts01(260 bp),ts02(330 bp),ts03(420 bp),ts04(600 bp)。4个差异cD-NA片段的RNA杂交结果显示,只有ts04存在明显差异,在盐胁迫条件下有杂交斑点而在对照中没有杂交斑点,其余3个cDNA片段在盐胁迫和对照中都没有杂交斑点。进一步将ts04克隆并进行DNA序列测定及同源性分析。结论ts04可能是耐盐相关cDNA片段,该片段核酸序列与麦属抗性相关的肌动蛋白基因有23%同源。     

mRNA差异显示PCR技术在水稻研究中的应用

mRNA差异显示PCR技术是在基因转录水平上研究基因差异表达和性状差异的有效方法之一,该方法在生物的发育、分化和对各种生物、理化因子作用时应答过程基因表达的研究中应用十分广泛。就mRNA差异显示PCR技术的基本原理、优点与不足、改进与完善等作了简要归纳,综述了该方法在水稻的发育分化、激素调控、抗逆性和抗病性等方面所取得的研究成果,并对该方法在水稻方面的应用前景作了简单分析。     

Novel gene expressed during early embryogenesis of zebrafish identified by mRNA differential display

Following the revelation of the molecular mechanism of morphogenesis in fruitfly, research on the molecular mechanism of morphogenesis in vertebrate becomes the focus of developmental biology. The isolation of genes controlling the embryogenesis of zebrafish, a vertebrate model animal, is considered as an initial step toward investigating this issue. There are several approaches that can be used to isolate developmental genes, each of which is suited to a particular situation. In this note, mRNA differential display was utilized to demonstrate the mRNA differences among zebrafish embryos at 4, 5 and 6 h post fertilization (28.5℃, corresponding to oblong, dome and shield stages, respectively, called blastula, gastrula and neurula in this note). One cDNA tag that was specific to embryos at neurula stage was cloned and sequenced. After sequence comparison in Genbank, we found that this cDNA tag represents a novel gene. The expression of this gene in the developing zebrafish embryos was examined by whole mount in situ hybridization. The hybridization results confirmed that this gene was specifically expressed in zebrafish neurula embryos.     

Studies of the energy release of mitochondria from sporophyte cytoplasmic male sterile rice by differential scanning calorimetry

The differential scanning calorimetric (DSC) curves of the mitochondria isolated from two varieties of sporophyte cytoplasmic male sterile and their fertile lines of Yie Bai and Ma Xie type rice have been determined. The curves show that the energy is released continuously as temperature rise to 70°C. Some thermodynamic and kinetic parameters of the energy release of the mitochondria have been obtained. The presented results showed that the mitochondria from cytoplasmic male sterile rice released more heat and they had higher energy barrier, less rate, and more complicated mechanism than that of their fertile lines in the energy release process. Foundation item: Supported by the National Natural Science Foundation of China, the China Postdoctoral Science Foundation Biography: Zhou Pei-jiang (1956-), male, PhD, Associate professor. Current resarch interest is in biophysical chemistry and environmental ecological chemistry     

Isolation of a vernalization-related cDNA clone (VRC) using mRNA differential display in winter wheat

Vernalization is an essential factor which affects the flowering development in cold-requiring plants. There is a key stage of nucleic acid and protein metabolism in the process of vernalization in winter wheat. To probe into the molecular determinants of vernalization, we examined mRNA populations in differently-treated plumules of winter wheat (Triticum aestivum L. cv Yanda 1817) using mRNA differential display. One vernalizationrelated cDNA clone (VRC), VRC54, was identified and was only expressed at the key stage of 20 d vernalization, rather than at other stages of nonvernalization, 4 d vernalization and devernalization. Northern blot and sequence analysis indicated that VRC54 was a novel vernalization-related clone found in higher plant which not only might play an important role in the floral induction in vernalization-requiring plants but also was different from the cold-acclimatized genes.     

Isolation of cDNA fragment of fertility-related gene in photoperiod-sensitive genic male sterile rice

The photoperiod_sensitive genic male sterile rice (PGMR) is particularly useful to take advantage of heterosis in rice. mRNA differential display was used to isolate the fertility_relative genes in rice. After establishing an optimized mRNA differential display system, one of the differential cDNA fragments that maybe related to the development and maturation of rice panicle was cloned from a PGMR Nongken 58S.     

mRNA differential display on gene expression in settlement metamorphosis process of Ruditapes philippinarum larvae

The mRNA differential display （DDRT-PCR） technique was adopted to find out the genes related to settlement metamorphosis development process of Ruditapes philippinarum larvae. In this study, we have obtained three hundred and forty-six amplification bands in total from pediveliger larvae, veliger larvae, eye spot larvae and post-larvae. Sixty-five out of three hundred and forty-six bands are distinctly differential display from band pattern, which can be put into four groups, standing for different expression characters. Sixteen differential display bands were cloned, sequenced and analyzed and nine different sequences are obtained in the study. Three sequences have higher similarity to the cDNAs deposited in database and three are very similar to the rDNA of other species, considered as the rDNA of Ruditapes philippi narum. The rest three sequences are found to be novel sequences after analyzed. Their accession numbers are AY916799, AY916798, and AY916797 respectively. We thought the novel sequences are possibly relevant to the early embryo development of Ruditapes philippinarum larvae and can provide some fundamental understandings that are helpful for the improvement of scallop seed raising industry.     

用差异显示技术分离空心莲子草抗旱相关基因

采用mRNA差异显示技术,分离空心莲子草在干旱胁迫1 d后差异表达的基因,获得139个基因片段.经过Reverse Northern检测,初步证实有两个片段受干旱诱导表达上调.对其中一个片断克隆测序并进行核甘酸序列比对,显示与其他基因同源性都很低,表明可能是新的基因.半定量PCR技术证实该基因受干旱诱导表达上调.     

Comparison of the community structure of planktonic bacteria in ballast water from entry ships and local sea water in Xiamen Port

In this study, the bacterial community structures in samples of ballast water collected from a ship from Singapore and of local sea water collected from Xiamen Port were compared using restriction fragment length polymorphism (RFLP) and 16S rDNA sequence analysis. Except for dominant α-Proteobacteria that are common to both systems, the bacterial community structures of the two systems were quite different. Most of the clones derived from the different systems were grouped into different phylogenetic clusters, and the sys-tems share only one common RFLP pattern. The ballast water, which is likely from clean offshore waters, contains sequences specific to α- and γ-Proteobacteria. Phylogenetic analysis revealed that the ballast water contained sequences belonging to attached bacteria and bacteria commonly found in the open sea, as well as many novel sequences. In addition, no known pathogenic bacteria were detected in the ballast water samples. Conversely, water samples from Xiamen Port were apparently affected by the near shore environments.Specifically, in addition to α- and γ-Proteobacteria, water from Xiamen Port contained β- and δ-Proteobacteria, Synechococcus, Bacter-oidetes and Actinobacteria, which are common in coastal environments. Additionally, four pathogenic bacterial sequences and one plas-mid sequence of a potential red tide forming alga were detected in the water from Xiamen Port, which suggests that the local sea water is polluted. The results of this study can be used as background information to assess the risk associated with the introduction of non-indig-enous species to local systems and to establish ballast water management systems.     

乙肝病毒前C基因变异对宿主T细胞亚群的影响及其临床意义的研究

目的　通过检测乙型肝炎病毒 (HepatitisBVirus,HBV)变异株感染患者外周血T细胞亚群分布的变化 ,探讨HBV基因组前C区 1896位基因变异与宿主机体免疫水平的关系 .方法　用酶联免疫吸附试验 (ELISA)检测 5 5名患者血清乙型肝炎病毒标志物 (HBVM)HBsAg ,抗 HBs,HBeAg ,抗HBe及抗 HBc;采用定性PCR法检测上述患者血清HBVDNA ;通过限制性片段长度多态性分析法 (RFLP)检测发生前C区 1896位基因突变的HBV变异株 ;采用流式细胞术 (flowcytometry)对上述患者外周血的CD4 + T细胞 ,CD8+ T细胞 ,CD3+ T细胞含量进行检测 ,将检测结果同所设健康对照组进行比较 ,并对数据进行统计学分析 .结果　在 5 5名患者中 ,HBsAg(+) ,anti HBs(- ) ,HBeAg(+) ,anti HBe(- ) ,anti HBc(+)者共 2 2例 ,其HBVDNA检测结果均为阳性 ,在这2 2例患者中 ,有 2 0例被检出为单纯HBV野毒株感染 ,另有 2例被检出为变异株和野毒株混合感染 ;HBsAg(+) ,anti HBs(- ) ,HBeAg(- ) ,anti HBe(+) ,anti HBc(+)者共 33例 ,其中HBVDNA阳性者为 18例 .在 18例HBeAg(- )而HBVDNA (+)的患者中 ,有 17例被检出为HBV前C区 1896位变异株感染 .变异株感染患者外周血CD4 + T细胞含量较之健康对照组有所减低 (P <0 .0 1) ,CD8+ T细胞含量较之健康对照组也有所     

Identification of genes induced during Medicago sativa nodule development by using the cDNA-AFLP technique

Under limited nitrogen conditions, rhizobia are ableto induce the formation of nitrogen-fixing root nodules on their leguminous plant host. This organogenetic process is triggered by a complex exchange of molecu- lar signals between the host plant and bac…     

一类非线性多延迟微分方程一般线性方法的稳定性

给出了一类非线性多延迟微分方程(简记为MDDEs)一般线性方法GAR(p,q)-稳定的一个充分条件.     

一类带p(t)-Laplacian算子的分数阶微分方程边值问题解的存在性

研究了一类带p(t)-Laplacian算子的分数阶微分方程边值问题,利用Schaefer不动点定理得到了解的存在性,并举例验证其主要结论.p-Laplacian算子是p(t)-Laplacian算子的特殊形式,所得结果推广和丰富了已有结果.     

Fluid inclusions evidence for differential exhumation of ultrahigh pressure metamorphic rocks in the Sulu terrane

Since the discovery of coesite and diamond inclusions in eclogites from the Dabie-Sulu orogen, east-central China[1―3], this largest ultrahigh pressure (UHP) metamor- phic terrane in the world has attracted extensive scientific interests. A number of hydrous minerals such as zoisite, phengite, magnesite and talc have been found in the UHP rocks, showing that fluids have played an important role in this type of extreme metamorphic evolution[4―8]. Sev-eral techniques have been applied to th…     

某类高阶线性微分方程解的导函数的不动点与超级

研究了几类整函数系数的高阶线性微分方程解的导函数的不动点的问题,得到：由于受到微分方程的制约,该类方程解的导函数的不动点密度与解的增长性有着密切联系。     

16S rDNA序列技术分析鉴定颗粒污泥中的微生物

采用16S rDNA序列分析技术对降解五氯酚的微氧颗粒污泥形成过程中真细菌和古细菌的种群多样性和动态变化进行了研究.通过对DGGE主要条带进行序列比对,发现颗粒污泥中真细菌和古细菌都与不可培养的微生物具有很高的相似性,微氧颗粒污泥中同时存在好氧菌、微氧菌和厌氧菌.通过比较不同菌的相对数量变化发现五氯酚驯化后的颗粒污泥中产生了一系列对五氯酚降解有利的优势细菌和古菌,如Proteobacteria、Sphingomonas、Methanogenic bacterium等.