Significance of consensus CYC‐binding sites found in the promoters of both ChCYC and ChRAD genes in Chirita heterotricha (Gesneriaceae)

Abstract CYC‐like genes are widely conserved in controlling floral dorsoventral asymmetry (zygomorphy) through persistent expression in corresponding domains in core eudicots. To understand how CYC‐like gene expression is maintained during flower development, we selected Chirita heterotricha as a material and isolated the promoter sequences of the ChCYC1C and ChCYC1D genes, homologs of CYC, by inverse polymerase chain reaction. Further promoter analyses led to the identification of a putative cis‐regulatory element in each promoter matching the consensus DNA binding site for Antirrhinum CYC protein: GGCCCCTC at ?165 for ChCYC1C, and GGCCCCCC at ?163 for ChCYC1D. This indicates that both the ChCYC1C and ChCYC1D genes have probably evolved autoregulatory loops to sustain their expression in developing flowers. We also isolated the coding and promoter sequences of the ChRAD gene, a homolog of Antirrhinum RAD. Promoter analysis showed that the ChRAD gene promoter also contained a putative CYC‐binding site (GGCCCAC at ?134). Therefore, ChRAD is likely a direct target of the ChCYC1 genes, which is similar to Antirrhinum RAD. These results imply that the establishment of floral zygomorphy in Chirita may have been achieved by the evolution of an autoregulatory loop for CYC‐like genes, which was probably accompanied by simultaneous co‐option of the RAD‐like gene into their regulatory network.     

Hypomania: a transcultural perspective

This study examined the transcultural robustness of a screening instrument for hypomania, the Hypomania Checklist-32, first revised version (HCL-32 R1). It was carried out in 2606 patients from twelve countries in five geographic regions (Northern, Southern and Eastern Europe, South America and East Asia). In addition, GAMIAN Europe contributed data from its members. Exploratory and confirmatory factor analyses were used to examine the transregional stability of the measurement properties of the HCL-32 R1, including the influence of sex and age as covariates. Across cultures, a two-factor structure was confirmed: the first factor (F1) reflected the more positive aspects of hypomania (being more active, elated, self-confident, and cogni-tively enhanced); the second factor (F2) reflected the more negative aspects (being irritable, impulsive, careless, more substance use). The measurement properties of the HCL-32 R1 were largely invariant across cultures. Only few items showed transcultural differences in their relation to hypomania as measured by the test. F2 was higher among men and in more severe manic syndromes; F1 was highest in North and East Europe and lowest in South America. The scores decreased slightly with age. The frequency of the 32 items showed remarkable similarities across geographic areas, with two excep-tions: South Europeans had lower symptom frequencies in general and East Europeans higher rates of substance use. These findings support the interna-tional applicability of the HCL-32 R1 as a screening instrument for hypomania.     

环境因素对短命植物抱茎独行菜抽薹开花节律的影响

以新疆十字花科典型早春短命植物抱茎独行菜(Lepidium perfoliatum L.)为材料,分别在不同环境、不同土壤基质及不同春化时间下栽培,以探讨环境因素对抱茎独行菜抽薹开花的影响。结果表明:抱茎独行菜种子在蛭石∶珍珠岩(3∶1)中的出苗率显著高于营养土和自然生境土壤,基质对抱茎独行菜植株是否抽薹无显著影响,但影响其抽薹的早晚及结实特性;人工4℃春化对3种不同栽培环境中于阳台生长植株的抽薹有明显促进作用,而对培养室及户外环境中栽培植株是否抽薹无显著影响;抱茎独行菜抽薹开花对光照和温度的响应最明显,光照时间由短变长与苗期一定时间的低温之间的相互作用是促使抱茎独行菜抽薹开花的关键因素。     

HPV-2E2蛋白(A338V)突变体对E2蛋白DNA结合能力增强的研究

HPV-2是引起皮肤寻常疣的常见HPV型别,病毒E2蛋白可抑制病毒早期启动子的活性。我们曾经报道来自一例巨大寻常疣患者的HPV-2突变E2蛋白对病毒早期启动子活性的抑制作用明显减弱,该E2蛋白在其C末端的DNA结合区域带有A338V的点突变。本研究利用原核表达系统表达纯化了突变E2(A338V)和HPV-2原毒株的羧基端和全长蛋白。电泳迁移率实验结果显示,E2蛋白可与带有E2蛋白特异性结合位点的寡核苷酸探针形成复合物,突变E2蛋白比原毒株E2蛋白的DNA结合能力强。这提示DNA结合能力的增强可能为E2蛋白对病毒启动子活性影响的分子基础,与患者出现罕见巨大寻常疣这一临床表型关联。     

中华鲎虫外部形态结构的初步观察

中华鲎虫体由头、躯干和尾3部分组成.头部具1对复眼和1个无节幼体眼,复眼后有1个颈感器.第1触角细小、棒状,第2触角退化.口器由上唇、大颚和2对小颚组成,上唇为头部的突起物,大颚和2对小颚为附肢,第2小颚的皮肤皱褶延长扩大形成头胸甲.雌体躯干部由36个体节组成,雄体39个体节,前30节具附肢,共57对.雌体躯干部后6个体节无附肢,雄体躯干部后9个体节无附肢.尾节分叉,细长多节,具刚毛,左右两根尾鞭的分节不对称.     

球形芽孢杆菌Mtx1蛋白和苏云金杆菌Cyt1Aa晶体蛋白的协同作用

采用常规的生物测定方法确定了纯化的球形芽孢杆菌(Bacillus sphaericus)的缺失信号肽的97kDa营养期杀蚊毒素(Mosquitocidal toxin 1,Mtx1)蛋白和苏云金芽孢杆菌(Bacillus thuringiensis)27.3kDa的Cyt1Aa晶体蛋白对致倦库蚊(Culex quinquefasciatus)幼虫的杀虫活性。结果表明Mtx1和Cyt1Aa不同比例的混合物对致倦库蚊的毒力比单独毒素蛋白高,经统计分析表明两毒素蛋白对目标蚊幼虫具有明显的协同作用。在LC98处理浓度下,Mtx1和Cyt1Aa按3∶1混合的混合物LT50值比单独Mtx1的提前了6.36h。表明Cyt1Aa和Mtx1对致倦库蚊具有协同毒杀作用,提高对目标蚊虫的毒力、缩短半致死时间。该结果为深入研究Mtx1和Cyt1Aa的杀蚊作用方式奠定了基础,同时为其在蚊虫防治中的应用提供了新的思路和方法。     

细极链格孢菌peaT2基因在毕赤酵母中的表达及蛋白功能确定

从细极链格孢菌表达文库获得阳性克隆子,序列分析表明,克隆的DNA片段中含有完整的开放阅读框架,将该基因命名为peaT2(GenBank登录号为EF212880)。用PCR法扩增peaT2基因的编码序列并亚克隆到毕赤酵母表达系统的表达载体pPIC9K上,得到重组质粒pPIC9K/peaT2。重组质粒经SacⅠ线性化后用电穿孔法导入到毕赤酵母(Pichia pastoris)GS115中,采用MD、G418-YPD平板和PCR法筛选Mut 表型,获得了分泌表达的重组毕赤酵母。随机挑取一菌株作为表达菌,用甲醇诱导PeaT2蛋白表达。SDS-PAGE及Western blot检测结果均表明PeaT2在毕赤酵母中成功地分泌表达。用peaT2基因的表达蛋白处理小麦种子,生物测定表明,表达蛋白能明显促进小麦的生长,具有蛋白激发子作用。     

绿绒蒿自然杂交种及其亲本cpDNA trnL-trnF基因的遗传学分析

对自然杂交种Meconopsis×cookei及其亲本红花绿绒蒿M.punicea和五脉绿绒蒿M.quintuplinervia的叶绿体DNAtrnL-trnF区进行了序列测定,所得序列的长度为957～961bp,其中M.×cookei的序列长度为960bp,红花绿绒蒿为961bp,五脉绿绒蒿为957bp。利用软件Clustal X对所得序列进行排序和碱基比较,排序后的序列长度为964bp,其中trnLintron为512bp,trnL3′exon为50bp,trnL-trn Fintergenic spacer(IGS)为361bp,还包括41bp的trnF5′端片段。整个trnL-trnF区序列共有25个变异位点,其中杂交种M.×cookei与红花绿绒蒿具有相同碱基的位点有21个(占84%),M.×cookei与五脉绿绒蒿具有相同碱基的位点仅有1个(占4%),余下3个位点(占12%)中,M.×cookei的碱基与两个亲本均不相同。分析结果表明,杂交种M.×cookei的叶绿体基因trnL-trnF来自红花绿绒蒿,根据质体细胞质遗传的规律,从而推测红花绿绒蒿为该杂交种的母本,五脉绿绒蒿为其父本。     

甘蓝型油菜小核糖核蛋白BnSmD1编码区全长cDNA 的克隆与表达分析

通过筛选野生型油菜(Pet33-10)与无花瓣油菜(Apet33-10)反向消减文库(SSH)和运用RACE-PCR技术, 获得了甘蓝型油菜小核糖核蛋白BnSmD1的全长编码区 cDNA (GenBank登陆号DQ298446)。该基因长484 bp, 含有一个长354 bp的阅读框。BnSmD1在N端拥有两个高度保守的结构域(Sm-1和Sm-2), 羧基端则含有一个RG重复序列。Northern blot表达结果显示: BnSmD1在甘蓝型油菜的各个组织均有表达, 但是它在早期花蕾中的表达明显高于同期的叶和茎。通过对BnSmD1在Apet33-10无花瓣品系与野生型有花瓣品系Pet33-10中各组织的表达差异进行比较, 发现该基因在Apet33-10的早期花蕾中表达明显下降。因此, BnSmD1可能对植物的早期花发育起到了重要的作用, 并很有可能影响花瓣的形成。     

日本血吸虫信号转导蛋白Sjwnt_4基因的克隆、表达及功能分析

由Wnt基因家族产物与其它相关基因产物构成的Wnt信号通路,是细胞发育和生长调节的一个关键途径,对动物的发育特别是生殖系统的发育起重要的调节作用。在人类和小鼠中,Wnt4蛋白是性腺分化过程中主要调节因子,在胚胎发育中起着关键作用。利用RACE技术从日本血吸虫19d童虫中首次扩增到一个Wnt家族基因,序列分析表明该基因的完整编码框含1311bp,编码436个氨基酸,理论分子量49.6kD。同源性分析结果表明,该基因的氨基酸序列具有典型Wnt家族蛋白特征,与日本三角涡虫、人Wnt4的氨基酸序列相似性分别达43％、37％,推测为血吸虫的Wnt4基因,命名为Sjwnt4（GenBank登陆号DQ643829）。实时定量PCR分析显示该基因在14d童虫、19d童虫、31d虫体、44d雌虫及44d雄虫中均有表达,其中19d童虫中的表达量明显高于其它发育阶段,44d雌虫中的表达量明显高于雄虫。构建了该基因的原核表达载体pGEX_4T_2_Sjwnt4,应用大肠杆菌系统进行了表达,表达蛋白以包涵体形式存在,Western印迹显示表达产物能被日本血吸虫成虫粗抗原免疫血清所识别。Sjwnt4基因及其表达产物的获得,为探索Wnt信号通路对血吸虫发育、生殖的调节提供了重要基础。