斑马鱼Cyclin C的cDNA克隆及其在发育过程中的表达

细胞周期蛋白C(cyclin C)与细胞周期依赖性蛋白激酶cdk8结合,通过磷酸化RNA 聚合酶 II 和 TFIIH调控转录.在脊椎动物胚胎发育过程中有关细胞周期蛋白C合子型转录激活机制尚知甚少.本研究克隆了斑马鱼细胞周期蛋白C基因,并用Northern杂交和整体原位杂交技术检测其在胚胎发育过程中的表达状态.结果显示,斑马鱼细胞周期蛋白C高度保守,与人细胞周期蛋白C有88％的蛋白序列同源;斑马鱼细胞周期蛋白C在母型期开始表达,其表达伴随中囊胚转换时期的合子型转录激活以及整个胚胎发育过程.     

文昌鱼Rbx1同源基因的克隆，进化分析和表达图式的研究

在青岛文昌鱼中首次克隆到一个RING box同源基因，对其进行了序列特征和系统进化学分析，并且研究了该基因的胚胎发育和成体表达图式。AmphiRbx1演绎的蛋白序列与已知其他无脊椎动物和脊椎动物的同源分子表现出很高的相似性。利用原位杂交和RT-PCR技术研究该基因的表达，结果显示AmphiRbx1在胚胎发育各期均有表达，特别是在卵裂期胚胎和有高速分裂细胞的组织中有很强的表达。实验结果表明，Rbx1基因在进化中相当保守，AmphiRbx1可能在细胞分裂的调节中发挥作用。     

文昌鱼Rbx1同源基因的克隆，进化分析和表达图式的研究（英文）

在青岛文昌鱼中首次克隆到一个RING box同源基因,对其进行了序列特征和系统进化学分析,并且研究了该基因的胚胎发育和成体表达图式 。AmphiRbx1演绎的蛋白序列与已知其他无脊椎动物和脊椎动物的同源分子表现出很高的相似性。利用原位杂交和RT PCR技术研究该基因的表达,结果显示AmphiRbx1在胚胎发育各期均有表达,特别是在 卵裂期胚胎和有高速分裂细胞的组织中有很强的表达。实验结果表明,Rbx1基因在进化中相当保守,AmphiRbx1可能在细胞分裂的调节中发挥作用。
     

Isolation and expression pattern analysis of novel ESTs from human fetal brain

In order to learn the mechanism of brain development and differentiation, 90 expressed sequence tags (ESTs) were isolated by differential display from the cerebrum and cerebellum of 13-week and 33-week fetal brains. After searching database, 74 of them represented novel genes, some of them were homologous to the brain development related genes. Using total cDNA probes, 79 of the ESTs and their expression differences in fetal brain were further characterized.     

柳树NAC基因的克隆与表达模式分析

【目的】研究柳树重要抗逆转录因子基因家族,为揭示柳树抗逆分子机制提供理论依据。【方法】以‘苏柳2345’的转录组测序数据为基础,克隆了柳树NAC家族,并分析其序列结构、组织表达特异性以及不同胁迫条件下的表达模式。【结果】克隆的两个柳树NAC转录因子基因分别命名为SlNAC1和SlNAC2。生物信息学分析表明:SlNAC1序列全长为1 126 bp,编码产物含343个氨基酸残基,为稳定的亲水蛋白,定位于细胞核;SlNAC2序列全长为1 139 bp,编码产物含291个氨基酸残基,为稳定的亲水蛋白,定位于叶绿体。两个基因均具有典型的NAM结构域及A、B、C、D、E 5个亚结构域,还包括共同的LPPG、YPNG 和DEE保守基序及NAC抑制结构域。系统进化树分析显示, SlNAC1基因与木薯亲缘关系最近,SlNAC2基因与茄子亲缘关系最近。定量PCR结果显示SlNAC1和SlNAC2均在叶片表达,根中没有表达,为叶片特异性转录因子。定量PCR结果显示SlNAC1基因在脱落酸(ABA)和赤霉素(GA)处理24 h后显著上调,SlNAC2基因在聚乙二醇(PEG)、ABA和GA胁迫后显著上调。【结论】柳树SlNAC1基因在非生物胁迫下表达较为稳定,受ABA和GA胁迫诱导;SlNAC2受干旱、ABA和GA胁迫诱导,受影响明显高于SlNAC1,不受乙烯剂(ETH)胁迫影响。推测SlNAC1和SlNAC2参与GA、ABA信号传导,可能不参与ETH的信号途径。     

唐古特白刺NtCBL1、NtCBL2基因克隆及表达分析

[目的]唐古特白刺(Nitraria tangutorum)是土壤荒漠化和盐碱化防治的先锋植物,对高盐和干旱有极强的适应能力,植物CBL基因作为钙离子感受器,在植物逆境应答及发育过程中具有重要功能.以盐生植物唐古特白刺为材料,开展唐古特白刺CBL基因克隆及表达分析,以深入了解唐古特白刺的抗逆分子机制.[方法]根据唐古特...     

基因表达数据的频繁模式挖掘算法

使用关联规则的方法来挖掘基因表达数据,在研究分析经典关联规则挖掘算法优缺点的基础上,立足于基因表达数据所呈现的新特征,提出一种基于BSC_tree的分段与运算基因表达数据频繁模式挖掘新算法BSC-AND.实验结果表明,与FP-growth和FIS算法相比,所提出的算法具有更低的时间复杂性.     

Characterization,gene cloning and expression of new xylanase XYNB with high specific activity

Extracellular xylanase XYNB from Streptomyces olivaeeoviridis A1 has been purified and characterized.The optimal pH value and temperature of XYNB for its activity are 5.2 and 60℃, respectively. The specific activity of XYNB is as high as 2869.78 U/mg. Metal cations, EDTA and SDS have no effects on enzyme activity of XYNB. The gene xynB coding mature protein of XYNB has been cloned by PCR. The forward oligonucleotide primer used in the PCR reaction was synthesized based on the N-terminal amino acid sequence of XYNB mature protein, and the reverse oligonucleotide primers are random oligonucleotide. The cloned gene xynB is 576 bp long and its G C content is 64.3%. The xynB encodes 191 amino acid residues, and the putative molecular weight of XYNB is 20.839 kD. The xynB has been expressed in E. coli, and the expressed xylanase has normal bioactivity.     

人血管生成素基因的克隆及表达

利用RI-PCR方法从培养的人黑色素瘤细胞系A375中扩增得到了人血管生成素cDNA片段,测序正确后克隆入表达载体pET-28a( )中并转化于E.coli BL21宿主菌中.经IPTG诱导,表达了N端融合6个组氨酸标签(6His-tag)的血管生成素融合蛋白.利用6His-tag与过渡态金属离子Ni2 高亲和力结合的性质,经镍柱纯化,获得了高纯度的血管生成素融合蛋白,为进一步研究其生物活性及应用奠定了基础.     

Cloning and expression pattern of a dehydrin-likeBDN1 gene from drought-tolerantBoea crassifolia Hemsl.

A 500-bp cDNA fragment was amplified via RT-PCR from drought-induced total RNA of the drought-tolerant B. crassifolia Hemsl. using primers based on the sequence of published dehydrin conserved region. By using 5′RACE, full-length coding region (1 148 bp) of BDN1 gene was produced. It is a new member of the dehydrin gene family. Southern analysis indicated that BDN1 is present in the B. crassifolia genome as a single-copy gene. Northern analysis revealed that its expression is inducible by drought and cold stresses as well as ABA application.     

杨小舟蛾MtroOBP1基因的克隆、序列分析及表达

【目的】气味结合蛋白(odorant binding proteins,OBPs)是一种嗅觉相关的蛋白,该蛋白参与大多数气味分子的识别过程,并与气味分子相结合。获得杨小舟蛾[Micromelalopha troglodyta (Graeser)]OBPs以明确其特性。【方法】选择羽化后健康的杨小舟蛾触角为模板,通过反转录聚合酶链反应(RT-PCR)技术克隆杨小舟蛾MtroOBP1基因,利用生物信息学分析软件研究其基因序列和蛋白结构,运用实时荧光定量PCR(qPCR)技术研究MtroOBP1的组织表达模式。【结果】利用RT-PCR技术从杨小舟蛾触角总RNA中扩增得到MtroOBP1基因(GenBank登录号:MN056510),序列分析表明,MtroOBP1开放阅读框为777 bp,一共编码了258个氨基酸残基,且翻译的氨基酸序列仅含有4个保守的半胱氨酸位点,表明得到的OBP基因的编码蛋白不属于典型气味结合蛋白家族,而是属于Minus-C家族。蛋白质理化性质预测显示:MtroOBP1蛋白的分子量为29 664.57 u,等电点为6.23,有18个潜在的磷酸化位点,没有明显的跨膜区,疏水指数为-2.011~3.078,有一个由19个氨基酸组成的信号肽,说明为分泌型蛋白。组织表达模式表明MtroOBP1在杨小舟蛾的各部位都表达,但在触角中表达量最高。【结论】首次克隆得到杨小舟蛾MtroOBP1基因,在触角中高表达,推测其蛋白具有运输气味分子的功能,在杨小舟蛾的嗅觉识别中起着至关重要的作用。     

藏猪NRAMP1基因的定量表达研究

采用半定量和定量PCR方法对藏猪的NRAMP1基因在脾脏等10个组织中的表达进行了研究.结果显示,该基因在所研究藏猪的组织中均得以表达,无组织表达特异性;定量PCR的结果证实该基因在不同组织中的表达量有很大的差异,其中,在脾脏中表达量最高,在肝脏中表达量最低,两者之间相差77倍,10个组织的平均表达量为3233拷贝/μg cDNA;NRAMP1基因在10个组织表达丰度从高到低的顺序为：脾脏、大脑、肺脏、回肠、肾脏、颌下淋巴结、结肠、心脏、肌肉和肝脏.     

Cloning and expression of ophc2, a new organphosphorus hydrolase gene

The amino acid sequences of N-terminal and internal peptide of OPHC2, purified from Pseudomonas pseudoalcaligenes strain C2-1 in our lab, are determined. The full-length organphosphorus hydrolase gene ophc2 is cloned by PCR using the degenerate primers designed according to the sequences and future inverse PCR. The ophc2 gene is 975 bp long with G C content of 63%, comprising one open reading frame encoding a polypeptide of 324 amino acids with a molecular weight of 36 kD. The nucleotide sequence of ophc2 shows low homologies with those organphosphorus hydrolase genes deposited in GenBank, one of which exhibits the highest homology of 46.4% with ophc2. The organphosphorus hydrolase protein expressed in E. coli bears normal bioactivity.     

Analysis of gene expression patterns with cDNA microarray during late stage of spermatogenesis in mice

The differentiation process of round spermatids to spermatozoa during the late stage of spermatogenesis is called spermiogenesis. To explore spermiogenesis-related genes, cDNA microarray was used to study expression patterns of 1176 genes in pachytene spermatocytes, round spermatids and elongating spermatids of Balb/c mice. The results showed that 208 genes were detected in all the three cell types. Most of them were down-regulated from pachytene spermatocytes to round spermatids and elongating spermatids. However, up-regulation of 7 genes expression in round spermatids and 3 genes in elongating spermatids were found. Expression of 7 differentially expressed genes in cDNA arrays was further confirmed by semi-quantitative RT-PCR study. The RT-PCR results indicated that the expression of 6 genes was consistent with that in cDNA arrays, only one gene did not show differential expression by RT-PCR. These results may provide important clues for studying of expression, regulation, and function of spermiogenesis-related genes.     

斑马鱼crip2基因的克隆、亚细胞定位及在胚胎发育早期的表达分析

斑马鱼广泛用于脊椎动物遗传和发育生物学的研究．为揭示包含具有LIM结构域的crip基因的功能,本文以斑马鱼为实验对象,克隆了斑马鱼的crip2基因,分别构建了crip2-Teasy 和crip2-egfp载体,从含有crip2基因的crip2-Teasy质粒上转录crip2 RNA 探针,并用全胚胎原位杂交法检测crip2在斑马鱼胚胎中的表达,用crip2-egfp对crip2进行亚细胞定位研究,发现crip2基因在斑马鱼胚胎早期没有表达,五体节开始在脊索特异表达;后期,主要局限于在血管、心脏和体节间肌肉,表明crip2的表达并不局限于内皮系统．在细胞内,crip2基因在细胞核和胞浆中均有表达．这些工作为进一步研究crip2基因的功能奠定了基础．     

基因表达数据的频繁闭合模式挖掘新算法

基因表达数据集与传统事务数据集相比呈现出新的特征,由于其项目数远远大于事务数,使得大量现有的基于项目枚举的频繁闭合模式挖掘算法不再适用.为此提出一种频繁闭合模式挖掘新算法TPclose,使用TP-树(tidset-prefix tree)保存项目的事务集信息.该算法将频繁闭合模式挖掘问题转换成频繁闭合事务集挖掘问题,采取自顶向下分而治之的事务搜索策略,并组合了高效的修剪技术和有效的优化技术.实验表明,TPclose算法普遍快于自底向上事务搜索算法RERⅡ,最高达2个数量级以上.     

薄壳山核桃CiAGL6基因的克隆、亚细胞定位及表达

研究薄壳山核桃MADS-box转录因子CiAGL6基因的特性、亚细胞定位及在雌花不同发育时期的表达水平,为揭示薄壳山核桃成花分子机理提供理论依据。