Preservation of genes involved in sterol metabolism in cholesterol auxotrophs: facts and hypotheses

Background

It is known that primary sequences of enzymes involved in sterol biosynthesis are well conserved in organisms that produce sterols de novo. However, we provide evidence for a preservation of the corresponding genes in two animals unable to synthesize cholesterol (auxotrophs): Drosophila melanogaster and Caenorhabditis elegans.

Principal Findings

We have been able to detect bona fide orthologs of several ERG genes in both organisms using a series of complementary approaches. We have detected strong sequence divergence between the orthologs of the nematode and of the fruitfly; they are also very divergent with respect to the orthologs in organisms able to synthesize sterols de novo (prototrophs). Interestingly, the orthologs in both the nematode and the fruitfly are still under selective pressure. It is possible that these genes, which are not involved in cholesterol synthesis anymore, have been recruited to perform different new functions. We propose a more parsimonious way to explain their accelerated evolution and subsequent stabilization. The products of ERG genes in prototrophs might be involved in several biological roles, in addition to sterol synthesis. In the case of the nematode and the fruitfly, the relevant genes would have lost their ancestral function in cholesterogenesis but would have retained the other function(s), which keep them under pressure.

Conclusions

By exploiting microarray data we have noticed a strong expressional correlation between the orthologs of ERG24 and ERG25 in D. melanogaster and genes encoding factors involved in intracellular protein trafficking and folding and with Start1 involved in ecdysteroid synthesis. These potential functional connections are worth being explored not only in Drosophila, but also in Caenorhabditis as well as in sterol prototrophs.     

The outcome of phagocytic cell division with infectious cargo depends on single phagosome formation

Given that macrophages can proliferate and that certain microbes survive inside phagocytic cells, the question arises as to the post-mitotic distribution of microbial cargo. Using macrophage-like cells we evaluated the post-mitotic distribution of intracellular Cryptococcus yeasts and polystyrene beads by comparing experimental data to a stochastic model. For beads, the post-mitotic distribution was that expected from chance alone. However, for yeast cells the post-mitotic distribution was unequal, implying preferential sorting to one daughter cell. This mechanism for unequal distribution was phagosomal fusion, which effectively reduced the intracellular particle number. Hence, post-mitotic intracellular particle distribution is stochastic, unless microbial and/or host factors promote unequal distribution into daughter cells. In our system unequal cargo distribution appeared to benefit the microbe by promoting host cell exocytosis. Post-mitotic infectious cargo distribution is a new parameter to consider in the study of intracellular pathogens since it could potentially define the outcome of phagocytic-microbial interactions.     

甘蔗花叶病毒3’末端基因的克隆及外壳蛋白序列分析比较

选取我国SCMV优势株系A株系的分离物SCMV-CA为材料,经过病毒和病毒RNA的提纯,反转录获得病毒cDNA,并克隆到载体pUC19的SmaI位点上,筛选得到多个重组质粒。选取其中一个克隆SCMV-CA54进行测序,得到一个全长为1296 bp的核苷酸序列。这段序列由一个长为1044 bp的开放阅读框架（ORF）和一个长279 bp的3’末端非编码区序列（3'-UTR）及poly(A)尾巴组成。这个ORF包括病毒完整的外壳蛋白（CP）及部分核内含体蛋白b（NIb）基因序列。将所得序列同已知SCMV亚组中各株系分离物的核苷酸和氨基酸进行同源性比较,结果表明该序列与其它株系分离物CP核苷酸序列的同源性介于63.7%～77.6%之间,氨基酸的同源性介于64%～89%之间。根据马铃薯Y病毒属的序列同源性划分标准,SCMV-CA与其它株系或分离物的同源性关系均介于种与株系划分标准之间。这是我国首次报道SCMVCP基因序列。     

地衣芽胞杆菌在植物病害生物防治中的应用

随着对地衣芽胞杆菌研究的不断深入和其杀虫、抗菌、生物降解等多种生物学活性的发现,地衣芽胞杆菌被认为是芽胞杆菌属中最具有生物防治应用价值的菌种之一。本文论述了地衣芽胞杆菌的生物学特性及其防治植物病害的四种主要作用机制,包括竞争、分泌抗菌物质、诱导植物系统抗性和促进植物生长;介绍了地衣芽胞杆菌在水稻、棉花、番茄、芒果、辣椒等多种大田作物和经济果蔬上的应用现状和生物防治效果;并讨论了其在生防应用过程中存在的主要问题,为今后地衣芽胞杆菌在生物防治方面的研究提供理论依据和应用参考。     

Crystal structures of GI.8 Boxer virus P dimers in complex with HBGAs,a novel evolutionary path selected by the Lewis epitope

Human noroviruses (huNoVs) recognize histo-blood group antigens (HBGAs) as attachment factors, in which genogroup (G) I and GII huNoVs use distinct binding interfaces. The genetic and evolutionary relationships of GII huNoVs under selection by the host HBGAs have been well elucidated via a number of structural studies; however, such relationships among GI NoVs remain less clear due to the fact that the structures of HBGA-binding interfaces of only three GI NoVs with similar binding profiles are known. In this study the crystal structures of the P dimers of a Lewis-binding strain, the GI.8 Boxer virus (BV) that does not bind the A and H antigens, in complex with the Lewis b (Le^b) and Le^y antigens, respectively, were determined and compared with those of the three previously known GI huNoVs, i.e. GI.1 Norwalk virus (NV), GI.2 FUV258 (FUV) and GI.7 TCH060 (TCH) that bind the A/H/Le antigens. The HBGA binding interface of BV is composed of a conserved central binding pocket (CBP) that interacts with the β-galactose of the precursor, and a well-developed Le epitope-binding site formed by five amino acids, including three consecutive residues from the long P-loop and one from the S-loop of the P1 subdomain, a feature that was not seen in the other GI NoVs. On the other hand, the H epitope/acetamido binding site observed in the other GI NoVs is greatly degenerated in BV. These data explain the evolutionary path of GI NoVs selected by the polymorphic human HBGAs. While the CBP is conserved, the regions surrounding the CBP are flexible, providing freedom for changes. The loss or degeneration of the H epitope/acetamido binding site and the reinforcement of the Le binding site of the GI.8 BV is a typical example of such change selected by the host Lewis epitope.     

水稻空间技术育种的研究

为探讨空间条件对水稻育种的影响,利用返回式卫星搭载水稻干种子,返地种植,考察分析了12个亲本品种后代的性状表现及遗传与变异情况。结果表明：空间条件诱变的后代性状具有多样性变异和多向性遗传,且能稳定遗传,并有超亲遗传现象,诱变后代具有变异类型多、变异频率高、特殊突变体、育种周期短等特点。为进一步开发利用空间技术育种,在研究实践的基础上,提出了程序化、规范化、可操作性强的水稻空间育种技术规范。 Abstract:In order to inquire into the influences of space conditions on rice breeding,the dry seeds of 12 rice varieties were carried by recoverable satellite.After recovery,these seeds were sowed in the yield,the inheritance and variation of the characters of their progenies were observed and analysed.The results showed that the characters of the progenies mutated under space conditions segregated and varied in many adpects and directions,and the segregated and varied characters were heritable.These progenies posoessed significant transgressive inheritance,a plenty of variation types.and variation.was characterized by short breeding period.The resulfs in this study indicated that space technology breeding could be developed and used as a new method of mutation breeding.     

人胰岛素基因在体内外表达的研究

利用定点突变人胰岛素基因,以脂质体载体和质粒不同比例形成的复合物进行体内外转染。体外转染鼠肝细胞,G418进行筛选,利用放免法测定培养液中胰岛素含量;体内通过肝门脉注射,测定模型鼠血糖及转染后7天时血液中胰岛素的含量,结果显示目的基因已转入肝细胞,且体内外转染都有一定量的成熟胰岛素表达,体外转染中质粒与脂质体比为1∶6转染后24h表达量最高为10.45μIU/ml,体内转染使模型鼠的糖尿病症状明显改善,血糖最高降幅达55%。 Abstract:The transfection of mutated human insulin gene was studied using the complex of different proportion of plasmid and Liposome.Hepatic cell was used as the target cell in vitro,isolation of Hepatic cell including insulin gene was carried out by G418,the expression level of insulin in medium was measured by RIA method.The portal vein was cannulated with therapeutic gene in vivo,the blood glucose of the model was regularly examined and the insulin level was detected on the seventh day after transfection.The results showed that the target gene was transferred into the hepatic cell,expression of mature insulin was detected both in vivo and in vitro,It reached the peak 10.45μIU/ml on the 24th hour after transfection with the proportion 1:6 of.plasmid and Liposome in hepatic cell.Diabetic sympotom of the model was improved after transgene,the blood glucose could decrease 55% at the most.     

Antisense Tiam1 Down-Regulates the Invasiveness of 95D Cells in Vitro

Invasion and metastasis are the main death causes oftumor patients, and aberrant expression of some genescontributes to tumor cell invasion and metastasis [1]. Tiam1was firstly identified as a gene amplified by insertedretrovirus which can confer metastat…     

Cloning and characterization of two cytoplasmic dynein intermediate chain genes in mouse and human

Cytoplasmic dynein is a large multisubunit microtubule-based motor protein, which mediates movement of numerous intracellular organelles. We report here the identification of the human homologue of cytoplasmic dynein intermediate chain 1 gene (DNCI1) located on human chromosome 7q21.3-q22.1. The mouse orthologue (Dnci1) was identified along with another highly related gene, Dnci2, and their RNA in situ expression patterns were examined during mouse embryogenesis. Dnci1 was found to have a highly restricted expression domain in the developing forebrain as well as the peripheral nervous system (PNS), while Dnci2 displayed a broad expression profile throughout the entire central nervous system and most of the PNS. A dynamic expression profile was also found for Dnci2 in the developing mouse limb bud. The data presented here provide a framework for the further analysis of the functional role of Dnci1 and Dnci2 in mouse and DNCI1 in human.     

几种鳄分子系统发生的探讨

百年来关于扬子鳄的分类学位置存在着很多争议,本文测得扬子鳄、暹罗鳄和湾鳄的mtDNAND4和Cytb基因,并从GenBank中获得密西西比鳄和海龟的ND4基因和Cytb基因相应片段.用ClustalX1.8进行对位排列,以海龟为外群构建分子进化系统树.结果显示:在鳄类动物中,扬子鳄与密西西比鳄的亲缘关系最近,两者ND4基因序列碱基差异为20.68%,而Cytb基因序列碱基差异为14.43%;但扬子鳄与密西西比与鳄的分类问题仍将有待进一步探讨。 Abstract:A portion of mitochondrial ND4 and cytochrome b gene of 3 species of erocodilian was sequenced.Additional homologous sequences of Alligator mississippinisis from GenBank were aligned together and compared with Cheloniamydas.The phylogenetic trees were reconstructed with the neighbor-joining (NJ) algorithm.Alligator sinensis ismore related to Alligator mississippinesis than others though the diversity of their mitochondrial ND4 gene reached 20.68% while Cyt b gene 14.43%.It needs further study to determine the phylogenetic relationships of Alligatorsinensis and Alligator mississi ppinisis.