四膜虫有性生殖过程中特异表达基因TCD3的序列分析

含有CHROMO(chromatin organization modifier)结构域的蛋白参与了染色质结构的调节.本研究从嗜热四膜虫(Tetrahymena thermophila)全基因数据库(www.ciliate.org)和基因表达数据库(tged.ihb.ac.cn)中筛选到有性生殖特异表达TCD3(Tetrahymena chromo domain-containing protein 3)基因.该基因ORF为1 005bp,大核染色体中基因全长1 241bp,含有两个内含子.5′-RACE和3′-RACE表明5′端非编码区(5′-UTR)长度为40bp,3′端非编码区(3′-UTR)长度为90bp.TCD3基因预测编码334个氨基酸构成,含有一个CHROMO结构域.CHROMO结构域聚类分析表明四膜虫Tcd3蛋白可能与线虫CeCDP功能类似,参与异染色质的形成和DNA序列删除.     

中华绒螯蟹卵巢新基因EJO6的全长cDNA克隆和序列分析

利用RACE技术从中华绒螫蟹卵巢获得了新基因EJO6(Eriocheir japonica ovary gene 6,EJO6)的全长cDNA序列(GenBarnk 检索号：AYl85922)。该cDNA序列长度为1250bp,开放阅读框为690bp,编码229个氨基酸。根据氨基酸序列计算的相对分子质量和等电点分别为24270和11．9。同时用生物信息学的方法对该基因的结构和功能进行了初步分析。     

费尔干猪毛菜液泡膜氢离子焦磷酸酶基因SfVP的克隆、序列分析及表达

植物液泡膜氢离子焦磷酸酶(Vacuolar H+-PPase,VP)能够酸化液泡并为液泡的次级转运系统提供能量,在植物耐盐性方面发挥着着重要作用。本研究采用RT-PCR结合RACE方法,从费尔干猪毛菜中(Salsola ferganica)克隆得到液泡膜质子焦磷酸酶基因SfVP,该基因的cDNA全长2738 bp,包含1个2301 bp的完整开放读码框(ORF),编码766个氨基酸的多肽。氨基酸序列分析表明其属于H+-转运无机焦磷酸酶超家族成员,具有典型的H+-PPase结构域,其疏水性较强,含有12个跨膜螺旋结构。SfVP序列比对显示具有H+-PPase的3个保守序列CS1、CS2和CS3,其中CS1中含有保守的底物催化位点,与烟草的NtVP氨基酸序列相似性为95.3%。半定量RT-PCR结果表明,SfVP的转录表达丰度随着600 mmol/L NaCl处理时间的增加而升高,处理12 h后显著增加并一直维持在较高水平。     

Cloning and characterization of vernalization-related gene (ver203F) cDNA 3′ end

Based on the cDNA fragment sequence of vernalization-related geneverc203 cloned by differential screening in our lab, the 5′ primer has been designed. The cDNA 3′ end ofver203 gene (1 197 bp) has been cloned by the RACE method. And it is identified by Northern blotting that its expression is special in vernalization treatment. After comparing the sequence in the nucleotide sequence databases of Genbank, EMBL and DDBJ, the gene has homology withHordeum vulgare jesmonate-induced protein gene. It is suggested that this gene might be related to the signal transduction mediated by jamonate.     

Cloning and characterization of vernalization-related gene (ver203F) cDNA 3' end

Based on the cDNA fragment sequence of vernalization-related gene verc203 cloned by differential screening in our lab, the 5' primer has been designed. The cDNA 3' end of ver203 gene (1 197 bp) has been cloned by the RACE method. And it is identified by Northern blotting that its expression is special in vernalization treatment. After comparing the sequence in the nucleotide sequence databases of Genbank, EMBL and DDBJ, the gene has homology with Hordeum vulgare jesmonate-induced protein gene. It is suggested that this gene might be related to the signal transduction mediated by jamonate.     

黄蜀葵查尔酮合成酶基因AmCHS克隆及序列分析

从黄蜀葵(Abelmoschus manihot) 花瓣中通过简并引物克隆得到查尔酮合成酶基因cDNA 核心序列,根据核心序列设计特异引物,再应用5′RACE 和3′RACE 技术分别扩增该序列的5′末端和3′末端序列,最后通过序列拼接获得黄蜀葵查尔酮合成酶基因（AmCHS）cDNA全序列.序列分析结果表明AmCHS cDNA编码区长1170 bp,编码389个氨基酸.其氨基酸序列与其它已知高等植物CHS基因具有很高的同源性,并且包含了CHS具有的活性位点和催化位点等保守性位点.     

杨树抗杨四瘿螨相关基因的表达分析

采用抑制性消减杂交(suppression subtractive hybridization,SSH)技术,在构建杨四瘿螨诱导差异表达的抑制消减cDNA文库的基础上,通过RTPCR技术对候选基因进行了验证,其中有6条差异表达候选基因在4个时间点表达量有明显的增强。运用RACE(Rapid Amplification of cDNA Ends)技术克隆获得了一段全长2 053 bp的cDNA克隆,命名为PtWRKY,编码588个氨基酸。