CDK5-mediated phosphorylation of CP190 may regulate locomotor activity in adult female Drosophila

正The three-dimensional organization of the genome plays a crucial role in regulating gene expression patterns in metazoans(Ong and Corces,2014).The nuclear architectural proteins are known to facilitate the formation of topological domains within the genome through mediating inter-and intra-chromosomal interactions.In vertebrate,CCCTC-binding factor(CTCF)is the main     

丙肝病毒融合抗原基因NS3-C定点整合入衣藻 叶绿体基因组的研究

用PCR方法从丙型肝炎病毒(HCV) cDNA文库中克隆了两段DNA片段,即HC基因组非结构NS3区抗原基因(约0.7 kb)和核心抗原C区抗原基因(约0.6 kb)的cDNA片段。在两段cDNA间加入连接肽Ser-Pro-Gly-Ser的密码子序列,构建成融合抗原基因NS3-C。将该融合基因与衣藻叶绿体基因atpA的启动子和rbcL基因的3'末端连接,得到丙肝病毒融合抗原基因NS3-C表达盒,再将该表达盒与选择标记基因aadA表达盒和衣藻叶绿体基因组同源片段连接,构建成衣藻叶绿体转化载体pSS6。基因枪法转化衣藻叶绿体,经壮观霉素筛选获得转化再生的单藻落,对转基因衣藻的PCR和Southern杂交分析表明,融合抗原基因NS3-C已整合到衣藻叶绿体基因组中。 Abstract:　Two DNA fragments encoding the nucleocapsid (C) region protein and the non-structural region 3 (NS3) protein of hepatitis C virus(HCV) were amplified from cDNA library by using PCR method. The 5' terminal of C cDNA fragment was linked up with the 3' terminal of NS3 cDNA fragment by a oligonucleotide linker Ser-Pro-Gly-Ser to form a chimeric gene NS3-C, which was placed under the control of the chloroplast atpA promoter and rbcL 3' region of Chlamydomonas reinhardtii to construct the chimeric gene NS3-C cassette. Then the NS3-C cassette was linked with selectable gene aadA cassette and the chloroplast homologous fragments of Chlamydomonas reinhardtii to generate transformation vector pSS6. Chloroplasts of Chlamydomonas reinhardtii were transformed by particle bombardment. Plastid transformants were selected by their resistance to 100 mg/L of spectinomycin. PCR and Southern hybridization analysis showed that the chimeric gene NS3-C had been integrated into chloroplast genome of Chlamydomonas reinhardtii.     

植物的功能基因组学研究进展 Progress in Functional Genomics in Plants

基因组研究计划包括以全基因组测序为目标的结构基因组学和以基因功能鉴定为目标的功能基因组学两方面的内容。目前基因功能鉴定的方法主要有：基因表达的系统分析(SAGE)、cDNA微阵列、DNA(基因)芯片、蛋白组技术以及基于转座子标签和T_DNA标签的反求遗传学技术等。本文对上述各种技术的优缺点以及它们在植物基因功能鉴定中的应用进行了综述。 Abstract: The genome projects comprise the structural genomics focusing on determining the complete sequences of the genome and the functional genomics focusing on elucidating the biological function of genes.The rapidly evolving tools for functional genomics research include Serial Analysis of Gene Expression (SAGE),cDNA microarray,DNA (or gene) chips,proteome project and the reverse genetics technique based on the well-established transposon tagging and T?DNA tagging systems.In this paper,the advantages and disadvantages of such techniques and application of these techniques in plant functional genomics research are reviewed and future prospective are also presented.     

Gene controlled by promoter--P_(TH4) depending on whiG of Streptomyces coelicolor

The downstream gene controlled by promoter--PTH4 which is related to Streptomycesdifferentiation was cloned, and its sequence was determined by the dideoxy chain termination method. The results indicated that the 1597 bp of DNA fragment conferred a complete open reading frame (ORF). In searches of databases, the deduced product of the ORF was not homologous with any known proteins; it may be a new protein. The function of the gene was studied using the strategy of gene disruption; the actinorhodin could not be produced when this gene was disrupted. Therefore, this gene may be related to actinorhodin biosynthesis in Streptomyces coelicolor, and the result also shows that this gene may play a role in multiple level regulation of differentiation genes in Streptomyces.     

植物的功能基因组学研究进展

基因组研究计划包括以全基因组测序为目标的结构基因组学和以基因功能鉴定为目标的功能基因组学两方面的内容。目前基因功能鉴定的方法主要有：基因表达的系统分析(SAGE)、cDNA微阵列、DNA(基因)芯片、蛋白组技术以及基于转座子标签和T_DNA标签的反求遗传学技术等。本文对上述各种技术的优缺点以及它们在植物基因功能鉴定中的应用进行了综述。 Abstract: The genome projects comprise the structural genomics focusing on determining the complete sequences of the genome and the functional genomics focusing on elucidating the biological function of genes.The rapidly evolving tools for functional genomics research include Serial Analysis of Gene Expression (SAGE),cDNA microarray,DNA (or gene) chips,proteome project and the reverse genetics technique based on the well-established transposon tagging and T?DNA tagging systems.In this paper,the advantages and disadvantages of such techniques and application of these techniques in plant functional genomics research are reviewed and future prospective are also presented.     

The revolution of the biology of the genome

Sequence data of entire eukaryotic genomes and their detailed comparison have provided new evidence on genome evolution. The major mechanisms involved in the increase of genome sizes are polyploidization and gene duplication.Subsequent gene silencing or mutations, preferentially in regulatory sequences of genes, modify the genome and permit the development of genes with new properties. Mechanisms such as lateral gene transfer, exon shuffling or the creation of new genes by transposition contribute to the evolution of a genome, but remain of relatively restricted relevance.Mechanisms to decrease genome sizes and, in particular, to remove specific DNA sequences, such as blocks of satellite DNAs, appear to involve the action of RNA interference (RNAi). RNAi mechanisms have been proven to be involved in chromatin packaging related with gene inactivation as well as in DNA excision during the macronucleus development in ciliates.     

黄鳝激素敏感性脂肪酶基因Hsl染色体原位杂交定位

动物脂肪组织中的甘油三酯在数量上是最重要的储存能源。Hsl基因所编码的激素敏感性脂肪酶是一种多功能酶。它通过催化水解储存在脂肪组织中的甘油三酯,以及卵巢、肾上腺、睾丸和胎盘中的胆固醇酯,在机体能量供应和类固醇生成作用中发挥重要作用。本研究以放射性同位素和地高辛标记重组质粒pBS中所含猪Hsl基因作为探针,分别与Pst Ⅰ酶切的黄鳝基因组总DNA和有丝分裂染色体标本进行Southern杂交和荧光原位杂交(FISH)。结果显示,Southern杂交呈现一条带,片段长度约为11.5kb。同时,应用FISH定位Hsl基因于黄鳝5号染色体,相对着丝粒距离为78.35±1.26。该定位结果与应用“特定染色体DNA池”定位黄鳝Hsl基因结果相符,且定位结果更为精细。表明在淡水鱼类黄鳝基因组中存在Hsl基因,另一方面也首次提供黄鳝5号染色体上FISH杂交信息,从而为增加黄鳝染色体组中已知的遗传标记和建立高精度遗传图谱奠定基础。 Abstract:Adipose tissue triacylglycerols are the quantitatively most important source of stored energy in animals.Hormone-sensitive lipase encoded by hormone-sensitive lipase gene (Hsl) is a multifunctional enzyme that catalyzes the hydrolysis of triacylglycerol stored in adipose tissue and cholesterol esters in the adrenals,ovaries,testes and macrophages.Using pig Hsl gene inserted into pBS labeled by the radioactive isotope and the digoxigenin as the probes respectively,one band,11.5kb,has been shown to hybridized with total DNA of rice field eel digested with Pst Ⅰby Southern blotting and Hsl gene has been assigned to metaphase chromosome 5,at the position of 78.35±1.26 from the c entromere in rice field eel by fluorescent in situ hybridization (FISH).The mapping results are corresponding to that of “specific-chromosomal DNA pool”obtained by chromosome microisolation used to map gene and the mapping result is more accurate.The results of the study further illustrate the importance of the presence of Hsl gene in rice field eel genome and provide the first FISH mapping data for rice field eel chromosome 5.The current studies would advance the addition of known genetic markers and the construction of high resolution genetic map in rice field eel genome.     

水平基因转移

简要介绍了水平基因转移（horizontal gene transfer,HGT）的基本概念以及进行转移的主要方式：由质粒或病毒等介导的水平基因转移和基因的“直接”水平转移。并结合基因组序列分析,重点阐述远缘生物之间发生的广泛的基因交流,以及其与进化、系统发育和基因工程生物安全性问题的探讨。 Abstract:In this paper the conception of horizontal gene transfer (HGT) was introduced,and main mode of HGT was also enumerated as follows:HGT by medium such as plasmid and virus etc.and the HGT without any medium.The transfer of genes from one species to another especially between remote species was emphasized by the information from genome sequencing.The problems about evolution phylogenies and safety of GEMs (gene engineered microorganisms) for HGT were discussed.     

Recombination and Heterologous Expression of Aliophycocyanin Gene in the Chloroplast of Chlamydomonas reinhardtii

Heterogeneous expression of multiple genes in the nucleus of transgenic plants requires theintroduction of an individual gene and the subsequent backcross to reconstitute multi-subunit proteins ormetabolic pathways.In order to accomplish the expression of multiple genes in a single transformationevent,we inserted both large and small subunits of allophycocyanin gene (apcA and apcB) into Chlamydomonasreinhardtii chloroplast expression vector,resulting in papc-S.The constructed vector was then introducedinto the chloroplast of C.reinhardtii by micro-particle bombardment.Polymerase chain reaction and Southernblot analysis revealed that the two genes had integrated into the chloroplast genome.Western blot andenzyme-linked immunosorbent assay showed that the two genes from the prokaryotic cyanobacteria couldbe correctly expressed in the chloroplasts of C.reinhardtii.The expressed foreign protein in transformantsaccounted for about 2%-3% of total soluble proteins.These findings pave the way to the reconstitution ofmulti-subunit proteins or metabolic pathways in transgenic C.reinhardtii chloroplasts in a single transformationevent.     

Evolution of double MutT/Nudix domain-containing proteins: similar domain architectures from independent gene duplication-fusion events

The MutT/Nudix superfamily proteins repair DNA damage and play a role in human health and disease. In this study, we examined two different cases of double MutT/Nudix domain-containing proteins from eukaryotes and prokaryotes. Firstly, these double domain proteins were discovered in Drosophila, but only single Nudix domain proteins were found in other animals. The phylogenetic tree was constructed based on the protein sequence of Nudix_N and Nudix_C from Drosophila, and Nudix from other animals. The phylogenetic analysis suggested that the double Nudix domain proteins might have undergone a gene duplication-speciation-fusion process. Secondly, two genes of the MutT family, DR0004 and DR0329, were fused by two mutT gene segments and formed double MutT domain protein genes in Deinococcus radiodurans. The evolutionary tree of bacterial MutT proteins suggested that the double MutT domain proteins in D. radiodurans probably resulted from a gene duplication-fusion event after speciation. Gene duplication-fusion is a basic and important gene innovation mechanism for the evolution of double MutT/Nudix domain proteins. Independent gene duplication-fusion events resuited in similar domain architectures of different double MutT/Nudix domain proteins.     

用RACE结合cDNA文库筛选的方法获取新的锌指蛋白基因

大多数有重要功能的蛋白质都含相应的由保守氨基酸顺序组成的功能结构域。本文首先根据蛋白质功能结构域保守氨基酸序列设计简并引物,用PCR方法扩增出基因EST序列,再利用改进的快速扩增cDNA末端（RACE）方法从cDNA文库中扩增出基因非同源部位,然后以非同源序列为探针,筛选cDNA文库。利用此方法成功地从人骨髓cDNA文库中克隆到几个编码锌指蛋白并代表原有EST的新的全长cDNA。这一策略也应适用于筛选编码具有其他序列保守性功能结构域蛋白的基因。 Abstract:Most of the important functionally proteins contain the corresponding function domains that consist of conserved amino acid sequences.The study provided a method to identify novel genes that encode proteins containing important functionally domains with conserved sequences.First,primers were designed according to the sequence of the cDNA library vector and the ESTs that have been obtained by reverse PCR and degenerate primers encoding Zinc finger domain.The cDNA library DNA was used as template for PCR amplification.The amplified fragment that contains nonhomologous sequences of the cDNA was inserted into pGEM-T easy vector.The fragment was recovered and used as a probe for screening the cDNA library.Several cDNAs with full length that encode proteins with Zinc finger domain and represent the original ESTs have been successfully cloned from a human bone marrow cDNA library.This strategy can also be used in screening genes that encode proteins containing differential function domains with conserved sequences.     

Sperm Mediated Transfer of Genes into Goldfish and Detection by Polymerase Chain Reaction

Goldfish sperms were mixed with eggs for fertilization after incubation with antifreeze protein gene(AFP)from ocean pout for 30 min.A number of embryos and 145 adult goldfish were obtained.DNA from adult goldfish and embryos was extracted separately.Results of the amplification by PCRand Southern blot molecular hybridization indicate the integration of exogenous antifreeze gene intothe genome of a part of the recipient goldfish.Of the 45 samples detected by PCR,twelve showedpositive reaction with distinct hybridization band.The positive rate was 26%.     

Conservation of ribosomal protein gene ordering in 16 complete genomes

The organization of ribosomal proteins in 16 prokaryotic genomes was studied as an example of comparative genome analyses of gene systems. Hypothetical ribosomal protein-containing operons were constructed. These operons also contained putative genes and other non-ribosomal genes. The correspondences among these genes across different organisms were clarified by sequence homology computations. In this way a cross tabulation of 70 ribosomal proteins genes was constructed. On average, these were organized into 9-14 operons in each genome. There were also 25 non-ribosomal or putative genes in these mainly ribosomal protein operons. Hence the table contains 95 genes in total. It was found that: (i) the conservation of the block of about 20 r-proteins in the L3 and L4 operons across almost the entire eubacteria and ar-chaebacteria is remarkable; (ii) some operons only belong to eubacteria or archaebacte-ria; (iii) although the ribosomal protein operons are highly conserved within domain, there are fine variat     

Enabling technology and core theory of synthetic biology

Synthetic biology provides a new paradigm for life science research(“build to learn”) and opens the future journey of biotechnology(“build to use”). Here, we discuss advances of various principles and technologies in the mainstream of the enabling technology of synthetic biology, including synthesis and assembly of a genome, DNA storage, gene editing, molecular evolution and de novo design of function proteins, cell and gene circuit engineering, cell-free synthetic biology, artificial intelligen...