大鼠肾脏组织cDNA文库的制备和葡萄糖转运蛋白cDNA克隆的筛选

 大鼠的肾脏组织经匀浆后,提取纯化总RNA和mRNA合成cDNA,进行甲基化,加人工接头,最后连入载体(λgt11)和外壳蛋白包装步骤制成肾脏组织的cDNA文库。插入载体的cDNA长度为0.5kb到8kb之间。经反复稀释,测定出该文库的效价为1.5×10~7pfu/mL。     

拟南芥AtPSK3基因的克隆及序列分析

根据拟南芥基因组数据库提供的信息,首次以特异引物经PCR技术克隆到拟南芥硫肽激素α的一个前体基因———AtPSK3,并对其进行了测序。序列分析表明,所获得的AtPSK3基因全长为505bp,含有一个内含子和两个没有3′或5′非转译区的外显子,与数据库提供的序列比较,同源性为100%。     

Succession of bacterial community composition over two consecutive years in two aquatic systems: a natural lake and a lake-reservoir

The succession in bacterial community composition was studied over two years in the epilimnion and hypolimnion of two freshwater systems: a natural lake (Pavin Lake) and a lake-reservoir (Sep Reservoir). The bacterial community composition was determined by cloning-sequencing of 16S rRNA and by terminal restriction fragment length polymorphism. Despite large hydrogeological differences, in the Sep Reservoir and Pavin Lake the dominant bacteria were from the same taxonomic divisions, particularly Actinobacteria and Betaproteobacteria. In both ecosystems, these major bacterial divisions showed temporal fluctuations that were much less marked than those occurring at a finer phylogenetic scale. Nutrient availability and mortality factors, the nature of which differed from one lake to another, covaried with the temporal variations in the bacterial community composition at all sampling depths, whereas factors related to seasonal forces (temperature and outflow for Sep Reservoir) seemed to account only for the variation of the hypolimnion bacterial community composition. No seasonal reproducibility in temporal evolution of bacterial community from one year to the next was observed.     

早期人胚胎cDNA文库构建及目的基因筛选

收集受精后３、４和５周龄药物流产胚胎,用改良一步法提取总ＲＮＡ,ｏｌｉｇｏ（ｄＴ）纤维素柱纯化ｍＲＮＡ,逆转录合成一链ｃＤＮＡ,完成二链ｃＤＮＡ的合成后,经碱变性电泳检测,合成ｃＤＮＡ的大小为０．４～９．０ｋｂ之间,且主要集中在１．０～２．０ｋｂ。除去多余的接头,收集大于４００ｂｐ的ｃＤＮＡ片段,与载体ｐＳＰＯＲＴ１和和γＺｉｐＬｏｘ连接,分别得到３、４、５周龄人胚胎质粒文加和噬菌体文库。另外,采     

HCV核心抗原基因的克隆和表达

用ＰＣＲ方法扩增人丙型肝炎病毒（ＨＣＶ）Ｃ抗原部分基因片段,将其克隆到一种新的表达载体ＰＱＥ中构成重组质粒ＰＱＥ－Ｃｏｒｅ短,转化大肠杆菌Ｍ１５株。含ＰＱＥ－Ｃｏｒｅ重组质粒的工程菌株以２ＹＴ中３７℃下培养加入ＩＰＴＧ诱导７小时,经１５％ＳＤＳ－ＰＡＧＥ凝胶电泳分离蛋白质,证明工程菌合成了大量改造的ＨＣＶ Ｃｏｒｅｎｈｊ ｒ     

Towards a theory of ecotone resilience: coastal vegetation on a salinity gradient

Ecotones represent locations where vegetation change is likely to occur as a result of climate and other environmental changes. Using a model of an ecotone vulnerable to such future changes, we estimated the resilience of the ecotone to disturbances. The specific ecotone is that between two different vegetation types, salinity-tolerant and salinity-intolerant, along a gradient in groundwater salinity. In the case studied, each vegetation type, through soil feedback loops, promoted local soil salinity levels that favor itself in competition with the other type. Bifurcation analysis was used to study the system of equations for the two vegetation types and soil salinity. Alternative stable equilibria, one for salinity-tolerant and one for salinity intolerant vegetation, were shown to exist over a region of the groundwater salinity gradient, bounded by two bifurcation points. This region was shown to depend sensitively on parameters such as the rate of upward infiltration of salinity from groundwater into the soil due to evaporation. We showed also that increasing diffusion rates of vegetation can lead to shrinkage of the range between the two bifurcation points. Sharp ecotones are typical of salt-tolerant vegetation (mangroves) near the coastline and salt-intolerant vegetation inland, even though the underlying elevation and groundwater salinity change very gradually. A disturbance such as an input of salinity to the soil from a storm surge could upset this stable boundary, leading to a regime shift of salinity-tolerant vegetation inland. We showed, however, that, for our model as least, a simple pulse disturbance would not be sufficient; the salinity would have to be held at a high level, as a ‘press’, for some time. The approach used here should be generalizable to study the resilience of a variety of ecotones to disturbances.     

柚木无性系愈伤组织诱导及植株再生

以柚木优良无性系71-14组培苗节间茎段为材料,MS 为基本培养基,采用正交设计对6-BA、IBA、TDZ、NAA 等4个生长调节剂各4水平进行愈伤组织诱导,并以最佳组合使用不同浓度的 TDZ 进行柚木愈伤组织再生.结果表明：TDZ 对形成具再生能力的致密型愈伤组织影响最大,低浓度水平的 TDZ 和6-BA 更易形成致密型愈伤组织;以愈伤组织大小、诱导率和致密型所占比例采用隶属函数法评定得出最优的愈伤组织诱导培养基为 MS＋0．9 mg·L-16-BA＋0．04 mg·L-1 IBA＋0．02 mg·L-1 TDZ＋0．8 mg·L-1 NAA,愈伤组织诱导率达80．78％、平均直径1．65 cm,获致密型愈伤组织83．0％;得出优化的再生培养基为 MS＋0．132 mg·L-1 TDZ,分化率为34．22％;初步建立了以茎段为外植体的柚木优良无性系71-14的再生体系,为柚木转基因技术的研究提供技术支撑.     

芒果SEPALAATA基因的克隆与表达分析

以芒果品种‘吕宋’(Mangifera indica L.Carabao)为试材,利用同源克隆和RACE技术从花序中获得了1个芒果SEPALAATA(SEP)基因cDNA全长,命名为MSEP1(GenBank中登录号为KP702299)。MSEP1基因的cDNA全长为921bp,包含一个长度为726bp开放阅读框,编码241个氨基酸,蛋白质相对分子质量为27.7kD,理论等电点为5.79。序列比对和系统进化树分析表明,MSEP1具有保守的MADS-box及半保守的K区,属于MADS-box家族的SEP亚家族。器官特异性表达分析表明,MSEP1基因在芒果根、茎中表达量较低,在叶片、花芽中表达量较高,而在花序中表达量最高。研究推测,MSEP1基因可能在芒果生殖生长中发挥重要作用。