Identification, Characterization, and Regulation of a Cluster of Genes Involved in Carbapenem Biosynthesis in Photorhabdus luminescens

The luminescent entomopathogenic bacterium Photorhabdus luminescens produces several yet-uncharacterized broad-spectrum antibiotics. We report the identification and characterization of a cluster of eight genes (named cpmA to cpmH) responsible for the production of a carbapenem-like antibiotic in strain TT01 of P. luminescens. The cpm cluster differs in several crucial aspects from other car operons. The level of cpm mRNA peaks during exponential phase and is regulated by a Rap/Hor homolog identified in the P. luminescens genome. Marker-exchange mutagenesis of this gene in the entomopathogen decreased antibiotic production. The luxS-like signaling mechanism of quorum sensing also plays a role in the regulation of the cpm operon. Indeed, luxS, which is involved in the production of a newly identified autoinducer, is responsible for repression of cpm gene expression at the end of the exponential growth phase. The importance of this carbapenem production in the ecology of P. luminescens is discussed.     

与木葡聚糖合成的糖基转移酶基因研究进展

半纤维素多糖木葡聚糖(XyG)存在于大多数植物的初生细胞壁中, 对细胞壁的结构组织和生长发育具有重要的调控作用。XyG在植物进化中存在结构的多样性。该文概述了参与XyG合成的糖基转移酶的最新研究进展, XyG合成需要多种糖基转移酶参与, 这些酶类很可能以蛋白酶复合体的形式存在并发挥作用, XyG的结构和组成的改变对植物的生长发育也产生影响。     

参与木葡聚糖合成的糖基转移酶基因研究进展

半纤维素多糖木葡聚糖(XyG)存在于大多数植物的初生细胞壁中, 对细胞壁的结构组织和生长发育具有重要的调控作用。XyG在植物进化中存在结构的多样性。该文概述了参与XyG合成的糖基转移酶的最新研究进展, XyG合成需要多种糖基转移酶参与, 这些酶类很可能以蛋白酶复合体的形式存在并发挥作用, XyG的结构和组成的改变对植物的生长发育也产生影响。     

Characterization of Two Polyketide Synthase Genes Involved in Zearalenone Biosynthesis in Gibberella zeae

Zearalenone, a mycotoxin produced by several Fusarium spp., is most commonly found as a contaminant in stored grain and has chronic estrogenic effects on mammals. Zearalenone is a polyketide derived from the sequential condensation of multiple acetate units by a polyketide synthase (PKS), but the genetics of its biosynthesis are not understood. We cloned two genes, designated ZEA1 and ZEA2, which encode polyketide synthases that participate in the biosynthesis of zearalenone by Gibberella zeae (anamorph Fusarium graminearum). Disruption of either gene resulted in the loss of zearalenone production under inducing conditions. ZEA1 and ZEA2 are transcribed divergently from a common promoter region. Quantitative PCR analysis of both PKS genes and six flanking genes supports the view that the two polyketide synthases make up the core biosynthetic unit for zearalenone biosynthesis. An appreciation of the genetics of zearalenone biosynthesis is needed to understand how zearalenone is synthesized under field conditions that result in the contamination of grain.     

Identification of Genes Involved in Flavonoid Biosynthesis in Sophora japonica Through Transcriptome Sequencing

Sophora japonica is a traditional Chinese medicinal ingredient that is widely used in the medicine, food, and industrial dye industries. Since flavonoids are the main components of S. japonica, studying the flavonoid composition and content of this plant is important. This study aimed to identify molecules involved in the flavonoid biosynthetic pathways in S. japonica. Deep sequencing was performed, and 85,877,352 clean reads were filtered from 86,095,152 raw reads. The clean reads were spliced to obtain 111,382 unigenes, which were then annotated with NR, GO, KEGG, eggNOG. Differential expression analysis and NR function prediction revealed 18 differentially expressed unigenes associated with 13 enzymes in flavonoid biosynthetic pathways. Our results reveal new insights on secondary metabolite biosynthesis‐related genes in S. japonica and enhance the potential applications of S. japonica in genetic engineering.     

人参植物皂苷生物合成相关新基因的筛选与鉴定

人参植物根进行的特定发育过程在药用次生物———人参皂苷生物合成和累积中发挥重要作用。为从人参根中分离出人参皂苷生物合成相关基因 ,采用抑制差减杂交技术 ,构建四年和一年生人参根组织mRNA群体间正向差减cDNA文库。对从差减文库中筛选的 4 0个阳性cDNA克隆进行酶切、PCR与逆向Northern斑点杂交鉴定、DNA测序以及核苷酸序列同源性比较。结果表明 ,获得的 6个差减克隆在GenBank/DDBJ/BMBL无对应的同源基因 ,代表新基因序列。与此同时 ,使用Northern印迹杂交验证及半定量RT PCR进一步确认 ,6个转录本为根发育阶段差异性表达基因。因而提示 ,它们可能在人参皂苷生物合成中发挥了重要作用。此外 ,在人参茎、叶与种子中亦能检测到上述基因转录本的表达。目前 ,6个新基因已被命名 ,在GenBank注册并获登录号 ,为克隆上述新基因cDNA全长序列及深入鉴定其在人参皂苷生物合成中的功能提供了重要实验依据。     

Large Scale Identification of Genes Involved in Cell Surface Biosynthesis and Architecture in Saccharomyces Cerevisiae

The sequenced yeast genome offers a unique resource for the analysis of eukaryotic cell function and enables genome-wide screens for genes involved in cellular processes. We have identified genes involved in cell surface assembly by screening transposon-mutagenized cells for altered sensitivity to calcofluor white, followed by supplementary screens to further characterize mutant phenotypes. The mutated genes were directly retrieved from genomic DNA and then matched uniquely to a gene in the yeast genome database. Eighty-two genes with apparent perturbation of the cell surface were identified, with mutations in 65 of them displaying at least one further cell surface phenotype in addition to their modified sensitivity to calcofluor. Fifty of these genes were previously known, 17 encoded proteins whose function could be anticipated through sequence homology or previously recognized phenotypes and 15 genes had no previously known phenotype.     

Identification and Characterization of Brassinosteroid Biosynthesis and Signaling Pathway Genes in Solanum tuberosum

Russian Journal of Plant Physiology - Brassinosteroids (BRs) are plant hormones which promote plant growth and development. Their biosynthetic pathway and signal transduction pathway have been well...     

昆虫脂肪酸合成通路关键基因的研究进展

脂肪酸对昆虫生长、发育、繁殖、信息交流起到重要的作用。主要介绍脂肪酸合成通路中的5个关键基因,乙酰辅酶A羧化酶基因(ACC)、脂肪酸合成酶基因(FAS)、超长链脂肪酸延伸酶基因(ELO)、去饱和酶基因(desat)及脂酰辅酶A还原酶基因(FAR)在昆虫中的研究进展。     

萝卜乙烯合成途径基因的鉴定及对胁迫的响应

该研究通过生物信息学方法,对萝卜(Raphanus sativus L.)乙烯合成途径关键结构基因(MAT、ACS和ACO)家族成员进行鉴定,并利用转录组数据和荧光定量方法探究其组织表达和对生物及非生物胁迫的响应。结果表明:(1)萝卜基因组中包含8个MAT、16个ACS和7个ACO基因,均可分为3个亚家族。(2)这些基因启动子中至少含有1个光响应顺式作用元件;除ACO1.1和ACO3外,其余基因启动子中均至少含有一种响应植物激素的顺式作用元件;多个基因启动子中含有响应生物及非生物胁迫的顺式作用元件。(3)转录组数据分析发现,萝卜所有MAT、ACS6.1和ACO2/3/4在叶片中均具较高的表达量;根癌农杆菌侵染诱导抗病萝卜的MAT2-4、ACS2/7.1/7.2和ACO1.1/1.2/4显著上调表达,而感病材料多个基因下调表达;铅、镉和铬胁迫均显著促进MAT4.2和ACO1.1/4的表达,但抑制MAT1和ACO5.1的表达;4℃低温显著抑制MAT2.1/2.2/4.1和ACO1.1/5.2的表达。(4)qRT-PCR分析表明,NaCl和PEG-6000均显著促进ACO5.1/5.2的表达,但抑制MAT4.1和ACO4的表达;果糖和蔗糖可能参与了萝卜对PEG-6000胁迫的响应。该结果为研究萝卜乙烯合成途径基因家族成员的功能奠定了基础。     

Expression and Characterization of CYP52 Genes Involved in the Biosynthesis of Sophorolipid and Alkane Metabolism from Starmerella bombicola

Three cytochrome P450 monooxygenase CYP52 gene family members were isolated from the sophorolipid-producing yeast Starmerella bombicola (former Candida bombicola), namely, CYP52E3, CYP52M1, and CYP52N1, and their open reading frames were cloned into the pYES2 vector for expression in Saccharomyces cerevisiae. The functions of the recombinant proteins were analyzed with a variety of alkane and fatty acid substrates using microsome proteins or a whole-cell system. CYP52M1 was found to oxidize C₁₆ to C₂₀ fatty acids preferentially. It converted oleic acid (C_18:1) more efficiently than stearic acid (C_18:0) and linoleic acid (C_18:2) and much more effectively than α-linolenic acid (C_18:3). No products were detected when C₁₀ to C₁₂ fatty acids were used as the substrates. Moreover, CYP52M1 hydroxylated fatty acids at their ω- and ω-1 positions. CYP52N1 oxidized C₁₄ to C₂₀ saturated and unsaturated fatty acids and preferentially oxidized palmitic acid, oleic acid, and linoleic acid. It only catalyzed ω-hydroxylation of fatty acids. Minor ω-hydroxylation activity against myristic acid, palmitic acid, palmitoleic acid, and oleic acid was shown for CYP52E3. Furthermore, the three P450s were coassayed with glucosyltransferase UGTA1. UGTA1 glycosylated all hydroxyl fatty acids generated by CYP52E3, CYP52M1, and CYP52N1. The transformation efficiency of fatty acids into glucolipids by CYP52M1/UGTA1 was much higher than those by CYP52N1/UGTA1 and CYP52E3/UGTA1. Taken together, CYP52M1 is demonstrated to be involved in the biosynthesis of sophorolipid, whereas CYP52E3 and CYP52N1 might be involved in alkane metabolism in S. bombicola but downstream of the initial oxidation steps.     

Identification and Isolation of Genes Involved in Poly(β-Hydroxybutyrate) Biosynthesis in Azospirillum brasilense and Characterization of a phbC Mutant

Like many other prokaryotes, rhizobacteria of the genus Azospirillum produce high levels of poly(β-hydroxybutyrate) (PHB) under suboptimal growth conditions. Utilization of PHB by bacteria under stress has been proposed as a mechanism that favors their compatible establishment in competitive environments, thus showing great potential for the improvement of bacterial inoculants for plants and soils. The three genes that are considered to be essential in the PHB biosynthetic pathway, phbA (β-ketothiolase), phbB (acetoacetyl coenzyme A reductase), and phbC (PHB synthase), were identified in Azospirillum brasilense strain Sp7, cloned, and sequenced. The phbA, -B, and -C genes were found to be linked together and located on the chromosome. An A. brasilense phbC mutant was obtained by insertion of a kanamycin resistance cassette within the phbC gene. No PHB production was detected in this mutant. The capability of the wild-type strain to endure starvation conditions was higher than that of the mutant strain. However, motility, cell aggregation, root adhesion, and exopolysaccharide (EPS) and capsular polysaccharide (CPS) production were higher in the phbC mutant strain than in the wild type.     

海藻糖代谢途径相关基因及生物工程

海藻糖(Trehalose)是一种由两个葡萄糖分子通过α,α-1,l糖苷键连接的非还原性双糖。最早的记录是在19世纪初期作为黑麦的麦角菌的一种成分而被描述,后来发现海藻糖广泛存在于微生物、动物和植物体内,特别是在那些能抗脱水作用的生物中起着重要作用。这些特殊生物具有在脱水条件下存活多年的性质,包括所谓的“复苏植物”(Selaginella lepidophylla)、某些咸水虾、线虫及面包酵母等。当它们体内99％的水分被去掉之后,仍保持着能在获水后迅速复活的能力^[1]。研究表明,海藻糖对于生物抗逆具有重要的保护作用。海藻糖的应用研究因此得到了人们的广泛关注和重视,目前海藻糖已被用作酶、其它蛋白、生物制品甚至移植器官的保护剂。海藻糖作为生物体对抗环境胁迫的重要应激保护物质,在不同生物中存在多种合成和分解代谢途径,相关基因已相继被克隆和分析。海藻糖合成、分解及其调控是生物抗逆的重要机制,其相关基因的研究也是海藻糖生物工程的重要基础。