中国莲PHT1家族成员的生物信息学分析

以植物PHT1家族蛋白为研究对象,基于中国莲(Nelumbo nucifera)全基因组数据库,利用生物信息学方法发掘中国莲可能存在的PHT1家族成员,并对中国莲PHT1家族成员进行序列比对、系统进化、保守序列、跨膜结构、二级结构和亚细胞定位等分析。结果表明,从中国莲全基因组数据库中筛选获得4个PHT1蛋白,分别命名为:Nn PHT1:1、NnPHT1:2、NnPHT1:3、NnPHT1:4。它们在植物PHT1家族中属于GroupⅠ,均含有保守序列:GGDYPLSATIxSE。该序列具有11~12个跨膜区域,且保守序列位于第4个跨膜区域内,均定位于细胞质膜,且在细胞内侧第6和第7个跨膜区域之间均含有一个较大的细胞胞质内环结构;磷酸化位点和糖基化位点的数量范围分别在33~50个和4~9个;二级结构中α-螺旋和无规卷曲结构的数量总和均高于70%。研究结果展示了中国莲的PHT1家族成员的序列基本信息和结构特点,为深入研究中国莲的该家族成员基因及其编码蛋白的结构功能提供理论依据。     

天津地区番茄褪绿病毒的分子检测与基因组部分序列分析

为了确定天津番茄产区是否发生了番茄褪绿病毒病,了解该病毒天津分离物的主要基因是否发生变异,对该地区发生的To CV进行了分子检测,并对该病毒的外壳蛋白(CP)和热激蛋白70类似物(Hsp70h)的核苷酸和氨基酸进行了序列分析。结果表明,天津分离物CP蛋白的核苷酸和氨基酸序列与已报道的具有代表性的不同国家的分离物相似性均在97.3%以上。Hsp70h蛋白的核酸和氨基酸序列与已报道的具有代表性的不同国家的分离物相似性均在97.2%以上。表明CP和Hsp70h蛋白是2个最保守的蛋白。这是首次在分子水平上证明番茄褪绿病毒在天津地区为害。     

玫烟色棒束孢几丁质酶基因上游调控序列克隆与结构预测分析

为探明玫烟色棒束孢几丁质酶基因Ifuchi1的表达调控机理，根据Ifuchi1基因组全长核苷酸序列，采用基因组步移方法，获得Ifuchi1的5′-上游区序列(总长为2 402 bp)。与cDNA 序列进行比对，其中含有2个内含子。TFSEARCH 1.3 软件分析转录因子结合位点的结果显示，该序列中没有明显的TATA和CAAT框，含有4个可能的碳调控因子的结合位点5＇>(g/c)YggRg<3＇和3个氮调控因子的结合位点-相隔很近的GATA。其中含有可能的葡萄糖抑制调控序列和压力反应元件。本研究结果为深入研究玫烟色棒束孢几丁质酶基因的表达调控机制及其分子机理奠定了一定基础。     

猪肝脏基因组DNA提取与纯化的研究

对血液中提取基因组DNA的方法加以改进,获得了一种提取猪肝脏组织中DNA的有效方法。试验表明,利用SDS裂解,氯仿、异戊醇抽提母猪肝脏中基因组DNA,得到了纯度较高的DNA。     

基因组重排技术在链霉菌育种应用的研究进展

文中介绍了基因组重排技术的原理,并着重介绍了其在链霉菌育种方面的常规方法及主要过程,同时提出了解决一些常规问题的方法.     

From parasite genomes to one healthy world: Are we having fun yet?

In 1990, the Human Genome Sequencing Project was established. This laid the ground work for an explosion of sequence data that has since followed. As a result of this effort, the first complete genome of an animal, Caenorhabditis elegans was published in 1998. The sequence of Drosophila melanogaster was made available in March, 2000 and in the following year, working drafts of the human genome were generated with the completed sequence (92%) being released in 2003. Recent advancements and next-generation technologies have made sequencing common place and have infiltrated every aspect of biological research, including parasitology. To date, sequencing of 32 apicomplexa and 24 nematode genomes are either in progress or near completion, and over 600k nematode EST and 200k apicomplexa EST submissions fill the databases. However, the winds have shifted and efforts are now refocusing on how best to store, mine and apply these data to problem solving. Herein we tend not to summarize existing X-omics datasets or present new technological advances that promise future benefits. Rather, the information to follow condenses up-to-date-applications of existing technologies to problem solving as it relates to parasite research. Advancements in non-parasite systems are also presented with the proviso that applications to parasite research are in the making.     

绿色木霉基因组DNA不同提取方法的比较研究

为获得提取绿色木霉Tr9701孢子培养物DNA的简便方法,选取8种方法(破壁酶消解法、超声破碎法、-80℃冻融法、高盐溶液法、尿素溶液法、CTAB法、改良CTAB法、试剂盒法)提取DNA,再通过对提取物进行比色定量和PCR测定评价各种提取方法的优弊。结果表明,改良CTAB法和试剂盒法提取DNA的效果最好,可用于绿色木霉的后续分子生物学研究。     

国际鳞翅目昆虫基因组筑波会议与中国家蚕基因组

首届国际鳞翅目昆虫基因组研讨会于 2 0 0 2年 9月在日本召开 ,来自 12个国家的 12 0名科学家就鳞翅目昆虫基因组的研究现状进行了广泛的交流 ,并正式成立了鳞翅目昆虫基因组国际筹划指导委员会 ,初步决定在2 0 0 4年前完成家蚕这一鳞翅目的模式昆虫的全基因组测序。我国作为世界蚕丝业中心 ,有关家蚕基因组的研究应如何对应 ?值得业内人士深思。     

The S RNA segment determines symptom differences on Tetragonia expansa between two Watermelon silver mottle virus isolates

The WS-Y isolate of Watermelon silver mottle virus (WSMoV) causes severe necrosis in Tetragonia expansa. To determine the RNA segment that induces symptoms, genome reassorants between WS-Y and an isolate causing mild mottle, WS-O, were generated. The origin of each segment in the reassortants was identified by RT-PCR and subsequent restriction enzyme analysis of the amplified fragments. Thirty genome reassortants were isolated from co-infected T. expansa plants. The reassortants with the S RNA segment of WS-Y caused severe necrosis, while those with the S RNA segment of WS-O caused a mild mottle; hence, the S RNA determined symptom expression. The incidence of reassortants was disproportional among genotypes. The most frequent genome reassortant possesses the L RNA of WS-Y, the M RNA of WS-O and the S RNA of WS-Y. A similar ratio of genotypes was found in isolates of local lesions on Chenopodium quinoa. These results strongly suggested that competition occurred independently between the individual RNA segments in a co-infected T. expansa plant, not between isolates.     

基于TRAP的长穗偃麦草SCAR标记的开发及应用

为了开发长穗偃麦草的特异标记,依据TRAP技术,利用56对固定引物与随机引物的组合对中国春-长穗偃麦草附加系等材料进行PCR扩增,共筛选出160条分布于长穗偃麦草1E-7E染色体的特异扩增片段。通过对104个片段的序列进行同源比对,选择与小麦无同源序列的区段设计138对引物,分别对中国春、长穗偃麦草、中国春-长穗偃麦草附加系进行PCR扩增,最终获得30个长穗偃麦草特异SCAR标记,发展标记的效率为53.6%。利用这些特异SCAR标记对硬粒小麦(AABB)与异源六倍体小麦(AABBEE)杂交产生的F2中38个单株进行扩增鉴定,9个单株仅附加了1条相同的E染色体,其余为多条E染色体或者无E染色体附加。这些SCAR标记在不同材料和世代表现出优越的特异性和稳定性,可用于检测小麦背景中附加的相关长穗偃麦草染色体。