Inhibition of furin-mediated cleavage activation of HIV-1 glycoprotein gp160.

The envelope glycoprotein of human immunodeficiency virus (HIV) initiates infection by mediating fusion of the viral envelope with the cell membrane. Fusion activity requires proteolytic cleavage of the gp160 protein into gp120 and gp41 at a site containing several arginine and lysine residues. Activation at basic cleavage sites is observed with many membrane proteins of cellular and viral origin. We have recently found that the enzyme activating the haemagglutinin of fowl plague virus (FPV), an avian influenza virus, is furin. Furin, a subtilisin-like eukaryotic endoprotease, has a substrate specificity for the consensus amino-acid sequence Arg-X-Lys/Arg-Arg at the cleavage site. We show here that the glycoprotein of HIV-1, which has the same protease recognition motif as the FPV haemagglutinin, is also activated by furin.     

Sequence analysis of a 7.3 kb Bam H Ⅰ genomic fragment of fowlpox virus strain 282E_4

A 7.3 kb BamH I genomic fragment from fowlpox virus (FPV) strain 282E4 has been sequenced. After comparison with the FPV sequences collected in Genbank, and the entire genomic sequence of vaccinia virus (VV) strain Compenhagen, a 11.2 kb BamH I fragment from FPV strain Munich HP-438 has been found homologous to the fragment. Every predicted open reading frame (ORF) within the fragment has been compared with the ORFs of W strain Copenhagen at the amino acid level. The signal peptide and transmembrane region of each peptide encoded by each corresponding ORF predicted in the fragment have been studied as well.     

Sequence analysis of a 7.3 kbBam H I genomic fragment of fowlpox virus strain 282E4

A 7.3 kb BamH I genomic fragment from fowlpox virus (FPV) strain 282E₄ has been sequenced. After comparison with the FPV sequences collected in Genbank, and the entire genomic sequence of vaccinia virus (VV) strain Compenhagen, a ll.2 kb BamH I fragment from FPV strain Munich HP-438 has been found homologous to the fragment. Every predicted open reading frame (ORF) within the fragment has been compared with the ORFs of VV strain Copenhagen at the amino acid level. The signal peptide and transmembrane region of each peptide encoded by each corresponding ORF predicted in the fragment have been studied as well.     

Sequence of reovirus haemagglutinin predicts a coiled-coil structure

The use of modern techniques has led to new insights into the molecular mechanisms of viral pathogenesis. Although the infectious process is quite complex, it is clear that one critical stage, the interaction of viral attachment proteins with cell-surface receptors, often has a major role in determining the pattern of infection. The mammalian reoviruses have served as useful models for understanding the molecular basis of viral pathogenesis. The mammalian reovirus haemagglutinin (sigma 1 protein), which is an outer capsid protein, has been shown to be a major factor in determining virus-host cell interactions. To further our understanding of the structure and function of the haemagglutinin, we have cloned a complementary DNA copy of the reovirus type 3 S1 double-stranded RNA gene which encodes the virus haemagglutinin and have sequenced the DNA complementary to the S1 gene. Analysis of the predicted amino-acid sequence of the virus haemagglutinin has allowed us to determine that the amino-terminal portion contains an alpha-helical coiled-coil structure and that the carboxy-terminal portion contains the receptor-interacting domains. Using this information, we propose here a model of how the reovirus haemagglutinin is attached to the virus particle.     

人Rab蛋白cDNA的克隆和表达

从人胎脑cDNA文库中克隆到一种新的Rab cDNA,全长920bp,以编码213个氨基酸残基,该蛋白预测的分子质量为24567u,等电点7．34,经同源比较,该cDNA与GenBank数据库中登录号为X14964的Rab蛋白有83％的相似性和76％的相同性,将该cDNA克隆到经改造的PBV220表达质粒,转化DH5a菌株诱导表达出该蛋白,取24种不同组织的总cDNA各100ng,用该基因序列设计引物作PCR,结果在胎肝组织中检测到有明显条带,表明该Rab基因相对在胎肝有高表达。     

Nucleotide sequence and formation of the transforming gene of a mouse sarcoma virus

The complete nucleotide sequence of the transforming gene of a mouse sarcoma virus has been determined. It codes for a protein of 374 amino acids. The nucleotide sequence of the junctions between a murine leukaemia virus and cellular sequences leading to the formation of the viral transforming gene have also been elucidated. The viral transforming sequence and its cellular homologue share an uninterrupted stretch of 1,159 nucleotides, with few base substitutions. The predicted amino acid sequence of the mouse sarcoma virus transforming gene was found to share considerable homology with the proposed amino acid sequence of the avian sarcoma virus oncogene (src) product.     

Nucleotide sequence of the gene coding for the major protein of hepatitis B virus surface antigen.

DNA extracted from hepatitis B virus Dane particles has been cloned in bacteria using a plasmid vector. A full-length clone has been examined by restriction endonuclease analysis, and the nucleotide sequence of an 892-base pair fragment from cloned hepatitis B viral DNA encoding the surface antigen gene is reported. The amino acid sequence deduced from the DNA indicates that the surface antigens is a protein consisting of 226 amino acids and with a molecular weight of 25,398. The portion of the gene coding for this protein apparently contains no intervening sequences.     

绿豆防御素基因的克隆、序列分析和植物表达载体的构建

从绿豆叶片提取总DNA中,通过PCR方法扩增出362bp的具有多种抗病、抗虫特性的绿豆防御素基因, 并将其克隆到pGM-T easy vector,酶切图谱及DNA 测序分析表明克隆的片段包含了完整的绿豆防御素基因的编码序列,与原序列同源性达到99.5%,蛋白质同源性达到100%.此基因编码的多肽由73个氨基酸组成,含有28个氨基酸的信号肽和8个半胱氨酸,可形成4个二硫键.我们用此基因构建了高效植物表达载体pBin438-LD.     

内蒙古额吉淖尔盐碱湖一株产视紫红质的极端嗜盐古菌的分离及其bop基因核心序列及最适生长条件的分析

从内蒙古额吉淖尔湖分离得到一株产视紫红质的极端嗜盐古菌,初步鉴定为属于Natrinema属;并扩增出了其bop基因,该bop基因序列已在Genbank中注册,编号为KF863690,分析了其核心序列。结果表明bop基因开放阅读框为303 bp,编码101个氨基酸。blastp结果表明该蛋白属于Bac-rhodopsin蛋白家族,具有典型的细菌视紫红质蛋白结构域。利用BLAST进行相似性分析发现bop编码蛋白氨基酸序列与Genbank中编号AAS87571.1、WP-007110705和AFB77278.1的视紫红质蛋白相似性达到98%以上。此结果表明视紫红质蛋白在细菌及古菌中结构上具有很高的保守性,与其质子泵功能紧密相关。最后分析了该菌株的最适生长条件。该菌株在分离培养基中适宜生长的NaCl浓度范围为1.0~3.0 mol/L,适宜生长pH为6.5~8.5,适宜生长镁离子浓度为0.4~0.8 mol/L。通过分析比较得出该嗜盐古生菌株生长最适NaCl盐浓度为1.5 mol/L,最适pH值为7.5,最适镁离子浓度为0.4 mol/L。该实验结果为今后进一步研究产视紫红质的极端嗜盐古生菌株的生理生化特征及遗传学特征奠定了基础。     

Ralstonia metallidurans CH34苯酚、甲苯降解基因的生物信息学分析

采用生物信息学方法对Ralstonia metallidurans CH34基因组序列进行了深入分析,推测了参与降解苯酚、甲苯/二甲苯降解的有关基因及功能,编码序列推测结果显示,R. metallidurans CH34中可能存在苯酚、甲苯/二甲苯间位降解代谢途径的基因操纵子及其调节因子和辅助因子,为后续进一步克隆和研究各基因的准确功能提供了有益的信息,奠定了基础.     

Nucleotide sequence of the rat skeletal muscle actin gene

The actins constitute a family of highly conserved proteins found in all eukaryotic cells. Their conservation through a very wide range of taxonomic groups and the existence of tissue-specific isoforms make the actin genes very interesting for the study of the evolution of genes and their controlling elements. On the basis of amino acid sequence data, at least six different mammalian actins have been identified (skeletal muscle, cardiac muscle, two smooth muscle actins and the cytoplasmic beta- and gamma-actins). Rat spleen DNA digested by the EcoRI restriction enzyme contains at least 12 different fragments with actin-like sequences but only one which hybridized, in very stringent conditions, with the skeletal muscle cloned cDNA probe. Here we describe the sequence of the actin gene in that fragment. The nucleotide sequence codes for two amino acids, Met-Cys, preceding the known N-terminal Asp of the mature protein. There are five small introns in the coding region and a large intron in the 5'-untranslated region. Comparison of the structure of the rat skeletal muscle actin gene with available data on actin genes from other organisms shows that while the sequenced actin genes from Drosophila and yeast have introns at different locations, introns located at codons specifying amino acids 41, 121, 204 and 267 have been preserved at least from the echinoderm to the vertebrates. A similar analysis has been done by Davidson. An intron at codon 150 is common to a plant actin gene and the skeletal muscle acting gene.     

内蒙古盐碱湖一株编码细菌视紫红质的嗜盐古菌bop基因全序列克隆及表达

细菌视紫红质(Bacteriorhodopsin,BR)是由bop基因编码的唯一膜蛋白,具有光推动质子泵的作用,为生命活动提供所需要的能量。为了研究高表达且具有生物活性的BR蛋白,应用PCR扩增技术,从内蒙古额吉淖尔盐碱湖分离得到的一株嗜盐古菌中扩增得到bop基因全序列;并构建了具有His6标签的原核表达载体p ET28a-IM-1,转化到大肠杆菌BL21(DE3),用IPTG诱导表达BR;并对表达产物进行SDS-PAGE鉴定分析。结果显示获得的bop基因全序列开放阅读框为672 bp,编码223个氨基酸,含有重组质粒p ET28a-IM-1的阳性菌株在IPTG诱导下高效表达了分子量约为25 k Da的BR蛋白。同源性分析表明该BR蛋白结构在古菌与细菌中具有相对较高的保守性,这与其质子泵功能密切相关。该BR蛋白为新发现的细菌视紫红质,其基因序列已递交Gen Bank,其登录号为KT873301。     

Primary structure of helix C to helix G of a new retinal protein in H.sp.xz515

The gene fragment encoding the retinal protein from helix C to helix G in a new strain of halobacteria, H.sp.xz515 has been amplified by PCR method. The nucleotide sequence of this fragment has been determined. The deduced amino acid sequence has been compared with halobium br and other two br-like proteins, ar-1 and ar-2. Results show that those amino acid residues in br, essential for proton pumping and binding to retinal, are conserved. The residue M145 in br may be important for isomerization reaction of retinal.     

黄角苔叶绿体rbcL基因编码区的序列分析

测定了黄角苔（Ｐｈａｅｏｃｅｒｏｓｌａｅｖｉｓ（Ｌ．）Ｐｒｏｓｋ）叶绿体ｒｂｃＬ基因编码区的１３９８个核苷酸序列,并且由此推导出对应的氨基酸序列．此基因中密码子的第２和第３位上Ａ／Ｔ所占的比例较高,分别为５３６％和７７５％．黄角苔ｒｂｃＬ基因的编码区与花旗松、菠菜、玉米和雷氏衣藻间在核苷酸序列上的同源性分别为８４０％,８１５％,７９３％和７６３％;在氨基酸序列上的同源性分别为９１．０％,８９．９％,８８．５％和８９．３％．为研究角苔植物的系统发育提供了核苷酸序列方面的资料．     

Single amino acid substitutions in influenza haemagglutinin change receptor binding specificity

The haemagglutinin (HA) glycoproteins of influenza virus membranes are responsible for binding viruses to cells by interacting with membrane receptor molecules which contain sialic acid (for review see ref. 1). This interaction is known to vary in detailed specificity for different influenza viruses (see, for example, refs 2-4) and we have attempted to identify the sialic acid binding site of the haemagglutinin by comparing the amino acid sequences of haemagglutinins with different binding specificities. We present here evidence that haemagglutinins which differ in recognizing either NeuAc alpha 2 leads to 3Gal- or NeuAc alpha 2 leads to 6Gal- linkages in glycoproteins also differ at amino acid 226 of HA1. This residue is located in a pocket on the distal tip of the molecule, an area previously proposed from considerations of the three-dimensional structure of the haemagglutinin to be involved in receptor binding.     

Cell-surface expression of influenza haemagglutinin from a cloned DNA copy of the RNA gene

By replacing either the eight early or the late genes of SV40 with a cloned copy of the influenza virus haemagglutinin gene we have constructed recombinant viruses which, in infected cells, express large quantities of haemagglutinin. This glycoprotein, over 10(8) molecules of which are produced per cell, is identical in molecular weight to authentic influenza virus haemagglutinin, accumulates at the cell surface and displays haemabsorbing activity.     

Cloning and expression analysis of MBLL cDNA

The mbl (muscleblind) gene of Drosophila encodes a nuclear protein which contains two Cys3His motifs. The mutation of mbl gene will disturb the differentiation of all the Drosophila's photoreceptors. Primers have been designed according to human EST086139, which is highly homologous to mbl gene. Human fetal brain cDNA library has been screened and a novel cDNA clone has been obtained. The 2595 bp cDNA, designated MBLL (muscleblind-like), contains an open reading frame which encodes 255 amino acids and has 4 Cys3His motifs (GenBank Acc. AF061261). The amino acids sequence shares high homology to Drosophila's mbl. The Northern blot and RNA dot blot hybridization of 43 human adult tissues and 7 fetal tissues show that MBLL is a widely expressed gene, but the expression amounts differ in these tissues.     

Cloning of a novel gene associated with human nasopharyngeal carcinoma

One EST N27741 with high expression in normal adult nasopharynx tissues but low expression in adult poorly differentiated squamous nasopharyngeal carcinoma has been selected out by the high-density cDNA array expression profiling technique. The differential expression has been confirmed by RT-PCR. One novel gene of 1096 bp has been cloned based on this EST. Bioinformatics analysis found that the new gene sequence contains a whole reading frame encoding 256 amino acids. There is a stop codon TAA in front of the 5′ end start codon, and a tailing signal AATAAA and poly A tail at the 3′ end. There is no homologous known gene found after searching by blasting this sequence to non-redundancy nucleotide database. Therefore it is considered a novel gene related to nasopharyngeal carcinoma.     

猫细小病毒 PCR 检测方法的建立及初步应用

摘要: 目的 建立猫细小病毒 PCR 检测方法,应用于猫临床样本中 FPV 的快速检测。方法 根据已发表的 FPV VP2 基因序列设计合成引物,并以此建立 FPV 的 PCR 检测方法,并对方法的特异性、敏感性、稳定性等进行验证。 用建立的方法对 33 份猫临床样品进行检测。结果 建立的 FPV PCR 检测方法与猫疱疹病毒Ⅰ型(FHV-1)、猫冠 状病毒(FeCV)、猫合胞体病毒(FeSFV)、猫免疫缺陷病毒(FIV)均无交叉反应;可检测病毒最小滴度为 5lgTCID50 / mL,相应的 DNA 模板浓度为 4. 9 × 102 拷贝/μL;FPV DNA 在 － 30℃冰箱放置 12 个月仍可检测出目的条带。应用 该方法从 33 份猫临床样本中检测出 21 份 FPV 核酸阳性。结论 建立的 FPV PCR 检测方法具有特异、敏感及稳定 的特点,适合于临床 FPV 的感染检测。