Differential expression of two distinct T-cell receptors during thymocyte development

The product of the T-cell receptor (TCR) gamma-gene has recently been found to be expressed on a subset of both peripheral cells and thymocytes. As an initial approach to understanding the role of this gamma-chain of TCR (TCR gamma) in T-cell development, we have studied the ontogeny of TCR expression at the protein level in the developing murine thymus. We show here that the first T3-associated TCR to be expressed in the developing thymus is a disulphide-linked heterodimer composed of a gamma-chain of relative molecular mass 35,000 (Mr 35K) and a 45K partner (termed TCR delta). This TCR gamma delta is first detected approximately two days before the appearance of cell-surface TCR alpha beta heterodimers. We report that N-glycosidase digestions reveal that all of the gamma-protein expressed on fetal thymocytes, as in adult CD4-8-(L3T4-, Lyt2-) thymocytes, bear N-linked carbohydrate side chains. The major gamma-gene transcribed in mature, alpha beta-bearing T cells (V gamma 1.2C gamma 2)encodes no N-linked glycosylation site so these results suggest that the fetal gamma delta receptor defines a distinct T-cell lineage whose development in the thymus precedes classical alpha beta-bearing cells.     

Coordinate expression of the murine Hox-5 complex homoeobox-containing genes during limb pattern formation

The homoebox-containing genes of the Hox-5 complex are expressed in different but overlapping domains in limbs during murine development. The more 5' the position of these genes in the complex, the later and more distal is their expression. Antero-posterior differences are also observed. A model is proposed that accounts for the establishment of these expression domains in relation to the existence of a morphogen released by the zone of polarizing activity. Comparison of these observations with the expression patterns of the genes of Hox complexes in the early embryo suggests that similar molecular mechanisms are involved in the positional signalling along the axes of both the embryonic trunk and the fetal limbs.     

基于非线性回归分析的差异基因选择方法

提出了一种新的用于微阵列基因差异表达多重假设检验的统计量计算方法,该方法利用基因表达值到各类样本数据中心的距离作为统计量进行多重假设检验,各统计量之间没有相关性,并且有效地减弱了数据噪声带来的假阳性结果,从而提高了多重假设检验的功效,所选择出的基因集也具有更好的分类能力.     

Retroviral transduction of T-cell antigen receptor beta-chain and myc genes

Support for multistage models of oncogenesis has been provided by several highly leukaemogenic retrovirus isolates that have transduced more than one host cell gene. Where functional studies have been performed, these retroviral oncogenes show synergy for in vitro transformation and leukaemogenesis. In naturally occurring feline leukaemias associated with feline leukaemia virus (FeLV), retroviral transduction of myc is a frequent oncogenic mechanism. But evidence suggesting that the FeLV v-myc genes might be insufficient for leukaemogenesis was provided by the latency (12 weeks) and clonality of FeLV/v-myc-induced tumours and the absence of demonstrable in vitro transformation by these viruses. In the search for secondary leukaemogenic events in FeLV/v-myc tumours, we have identified a case of FeLV transduction of a T-cell antigen receptor beta-chain gene. The proviruses carrying this gene (which we have named v-tcr) were a separate population from those carrying v-myc. In its normal role, the T-cell receptor beta-chain forms part of a multimeric complex involved in antigen recognition and T-cell activation. We suggest that v-tcr is a novel viral oncogene which assisted v-myc in the genesis of a naturally occurring case of thymic lymphosarcoma.     

黑莓果实发育过程中蔗糖磷酸合成酶基因的表达分析

【目的】蔗糖磷酸合成酶(sucrose phosphate synthase, SPS)是调控植物蔗糖代谢合成的关键酶,在植物光合产物的积累与分配方面有重要作用。本研究旨在探讨黑莓3个SPS基因的系统发育关系、编码的蛋白特性、在不同发育时期、不同组织中的时空表达特性,并分析其与黑莓发育的关系。【方法】以黑莓栽培品种‘宝森’(‘Boysenberry’)为试材,从中克隆和鉴定了3个 SPS 基因家族成员,利用生物信息学和荧光定量聚合酶链式反应(qRT-PCR)等方法对3个黑莓SPS 基因RuSPS1、RuSPS2和RuSPS3的氨基酸序列、保守作用元件、编码的蛋白特性、蛋白结构及进化关系进行分析,并对这3个基因在黑莓中的时空表达情况与酶活性进行了相关性分析。【结果】多重氨基酸序列比对显示,黑莓SPS蛋白具有植物SPS家族特有的2个保守蛋白结构域及2个相对保守的蛋白磷酸位点;系统进化分析表明,RuSPS基因分为A、B两个亚族,其中RuSPS1和RuSPS3为A亚族成员,RuSPS2为B亚族成员;保守作用元件分析表明, 除RuSPS2含基本的蛋白保守元件外, RuSPS1和RuSPS3都存在不同程度的片段缺失;序列分析和比较揭示了黑莓SPS基因与其他家族的不同特征。qRT-PCR分析显示,3个RuSPS基因在黑莓各个组织器官中均有表达,其中RuSPS1在叶片和果实中表达量较高,在花中的表达量较低;RuSPS2在发育成熟的果实中有大量的表达,在其他器官中表达量较低;RuSPS3在各器官中的表达均较高,说明 SPS基因表达具有明显的组织特异性,3个RuSPS基因都随着果实发育进程在果实和叶片中表现了表达增加的趋势。果实和叶片中SPS酶活性的变化与RuSPS基因表达水平一致。相关分析表明,叶片中SPS活性与RuSPS2显著正相关(P<0.05),果实中SPS活性与RuSPS1显著负相关(P<0.05)。【结论】3个RuSPS基因与黑莓果实发育过程中的蔗糖合成与代谢关系密切,均参与了黑莓的生长发育调控,其中叶片中SPS活性的变化一定程度上是由RuSPS2调控,果实中SPS活性的变化则是由RuSPS1调控。     

Selective immortalization of murine macrophages from fresh bone marrow by a raf/myc recombinant murine retrovirus

Myeloid precursors can be grown in vitro in the presence of specific growth factors; however, their expansion is limited by a competing process of terminal differentiation. Proto-oncogenes seem to be involved in cellular proliferation and/or differentiation and may also play a role in the myelopoietic process. Murine myeloid precursors which are grown in vitro with growth factors respond with augmented self-renewal upon infection with recombinant retroviruses carrying the v-myc or v-src oncogenes, suggesting a synergism or complementation between some viral oncogenes (v-onc) and certain growth factors. We now show that the combination of two v-onc genes (raf and myc) induces the selective proliferation of monocytic cells from fresh murine bone marrow (BM) in the absence of a specific growth factor supplement. Depending on the culture conditions these cells can either differentiate and cease to proliferate or grow continuously, thus mimicking the alternative pathways that can be followed by committed BM stem cells in vivo.     

菠菜SoSWEET蛋白家族基因鉴定与表达分析

以现有菠菜基因组信息为基础,通过生物信息学方法筛选鉴定16个菠菜SoSWEET蛋白家族成员,命名为SoSWEET1~SoSWEET16.氨基酸残基数量在648~1 140之间,分子质量在54 070.32~95 868.64 u之间,理论等电点（pI）在5.06~5.19之间.亚细胞定位预测有6个SoSWEET蛋白定位于细胞膜,5个SoSWEET蛋白定位于内质网,5个SoSWEET蛋白定位于细胞膜、内质网.系统进化分析将菠菜SoSWEET蛋白家族分成4个亚族,在此基础上对基因结构、保守基序、顺式作用调控元件等进行分析.共鉴定了10个高度保守基序,其中所有菠菜SoSWEET蛋白都包含基序1,2和4,是构成菠菜SoSWEET蛋白中最高度保守的部分.所有菠菜SoSWEET蛋白家族成员都含MtN3_slv和PQ-loop superfamily结构域.大多数SoSWEET蛋白家族的基因含有5个内含子.顺式作用元件预测结果表明,菠菜SoSWEET基因启动子上包含光响应、生长发育、植物激素响应和逆境胁迫响应等顺式作用元件.组织表达分析表明,所有SoSWEET基因在根、茎、叶和叶柄中都有表达,霜霉病胁迫处理后16个基因表现出不同响应变化.本研究为后续深入研究菠菜SoSWEET蛋白家族成员的功能提供了重要参考.     

拟南芥PAL基因家族鉴定及表达模式分析

为研究苯丙氨酸解氨酶（PAL）作为苯丙烷类代谢途径的关键酶,在植物响应环境胁迫过程中的重要作用,本研究利用HMMER、Pfam与SMART等工具,从拟南芥全基因组中筛选获得 9 个PAL成员,分析其编码酶蛋白的理化性质、二级结构与保守基序等信息.同时,结合转录组与定量PCR分析PAL成员在干旱与盐胁迫条件下的表达模式.结果表明,拟南芥PAL成员的二级结构主要由α-螺旋和无规则卷曲组成;亮氨酸和丙氨酸为含量较高的氨基酸残基;拟南芥PAL成员含有 20 种不同的保守基序,其中motif6含有ASG活性位点,为所有PAL成员所共有;较多PAL成员具有光响应、激素响应、胁迫响应和生长发育相关的顺式调控元件.转录组学与定量PCR分析发现,AT3G53260.1、AT3G53260.2与AT3G47660.1的mRNA表达水平在干旱与盐胁迫后发生显著变化,推测这 3 条基因可能参与拟南芥对干旱及盐胁迫的响应过程,表明PAL基因在调控植物生长发育与非生物胁迫响应过程中发挥重要作用.     

Diversity of murine gamma genes and expression in fetal and adult T lymphocytes

The search for the genes encoding the T-cell receptor alpha and chains revealed a third gene, T gamma (ref. 1), which shares with t T alpha (refs 2-7) and T beta (refs 8-15) genes a number of structure features, including somatic rearrangement during T-cell development. T gamma gene expression appears to be unnecessary in son mature T cells and is at its greatest in fetal thymocytes encouraging speculation that T gamma has a role in T-cell development and may be involved in the recognition of polymorphic major histocompatibility complex (MHC) products during thymic education. One argument against the participation of T gamma in such a process has been its apparently limited diversity, due to the small number of gene segments available for rearrangement. We here describe the identification of additional T gamma V-gene segments and demonstrate that they can be rearranged to previously identified J- and C-gene segments and are expressed in fetal thymocytes. In addition we describe a variety of patterns of T gamma mRNA processing which may be significant for T gamma gene regulation.     

棉花腺体形成相关的miRNA差异表达研究

植物miRNAs在基因表达、生长、发育等方面有十分重要的作用．本实验以棉花显性无腺体近等基因系为材料,利用miRNA基因芯片杂交技术,分析与腺体形成相关的miRNA的差异表达．结果表明,棉花腺体形成期共有30个miRNA有差异表达,其中表达上调的miRNA有24个,分别属于miR156,miR157,miR166和miR390家族．表达下调的miRNA有6个,分别属于miR149,miR169,miR289,miR705,miR1224和miR1227家族．miRNA家族作用的靶基因分析发现主要分布在发育     

Assessing the redundancy of MADS-box genes during carpel and ovule development

Carpels are essential for sexual plant reproduction because they house the ovules and subsequently develop into fruits that protect, nourish and ultimately disperse the seeds. The AGAMOUS (AG) gene is necessary for plant sexual reproduction because stamens and carpels are absent from ag mutant flowers. However, the fact that sepals are converted into carpelloid organs in certain mutant backgrounds even in the absence of AG activity indicates that an AG-independent carpel-development pathway exists. AG is a member of a monophyletic clade of MADS-box genes that includes SHATTERPROOF1 (SHP1), SHP2 and SEEDSTICK (STK), indicating that these four genes might share partly redundant activities. Here we show that the SHP genes are responsible for AG-independent carpel development. We also show that the STK gene is required for normal development of the funiculus, an umbilical-cord-like structure that connects the developing seed to the fruit, and for dispersal of the seeds when the fruit matures. We further show that all four members of the AG clade are required for specifying the identity of ovules, the landmark invention during the course of vascular plant evolution that enabled seed plants to become the most successful group of land plants.     

Scriptaid affects histone acetylation and the expression of development-related genes at different stages of porcine somatic cell nuclear transfer embryo during early development

Although the somatic cell nuclear transfer(SCNT) technique has been used extensively for cloning and generating transgenic pigs,the cloning efficiency is still very low.It has been proposed that the low efficiency of this technique is the result of incomplete epigenetic reprogramming and abnormal gene expression during early embryonic development.In this study,we investigate the effect of Scriptaid,a low-toxicity histone deacetylase inhibitor,on the developmental competence of porcine SCNT embryos.We found that treating SCNT embryos with 500 nmol/L Scriptaid for 15 h after activation significantly enhanced the blastocyst formation rate(27.7%) compared with the untreated group(control)(12.2%,P<0.05).Using an immunofluorescence technique to measure the average fluorescence intensity,we also found that treating SCNT embryos with Scriptaid increased the level of histone acetylation on histone H3 at lysine 14(acH3K14).Furthermore,treating embryos with Scriptaid increased the expression level of three genes that play important roles during embryonic development(Oct4,Klf4 at the blastocyst stage and Nanog at the 4-cell stage).Moreover,the expression level of the apoptosis-related gene Caspase-3 was significantly lower in the Scriptaid-treated SCNT embryos compared with the control SCNT embryos at the 4-cell and blastocyst stages.In conclusion,these results indicate that Scriptaid treatment improves the development and nuclear reprogramming of porcine SCNT embryos.