An avirulence gene,AvrLmJ1, from the blackleg fungus,Leptosphaeria maculans,confers avirulence to Brassica juncea cultivars

The fungus Leptosphaeria maculans causes blackleg of Brassica species. Here, we report the mapping and subsequent cloning of an avirulence gene from L. maculans. This gene, termed AvrLmJ1, confers avirulence towards all three Brassica juncea cultivars tested. Analysis of RNA‐seq data showed that AvrLmJ1 is housed in a region of the L. maculans genome which contains only one gene that is highly expressed in planta. The closest genes are 57 and 33 kb away and, like other avirulence genes of L. maculans, AvrLmJ1 is located within an AT‐rich, gene‐poor region of the genome. The encoded protein is 141 amino acids, has a predicted signal peptide and is cysteine rich. Two virulent isolates contain a premature stop codon in AvrLmJ1. Complementation of an isolate that forms cotyledonary lesions on B. juncea with the wild‐type allele of AvrLmJ1 confers avirulence towards all three B. juncea cultivars tested, suggesting that the gene may confer species‐specific avirulence activity.     

Functional complementation between the two homologous genes, ops and tps, during differentiation of Myxococcus xanthus

Summary Protein S is a development-specific protein of Myxococcus xanthus encoded by the tps gene. It has been shown that there are two extensively homologous genes (ops and tps) tandemly repeated in the same direction with a 1.4 kb spacer fragment between them (Inouye et al. 1983). Seven deletion mutants were constructed by removing the ops gene, the tps gene, segments of the spacer sequence or combinations of these regions. The deleted regions were replaced with DNA fragments carrying the Tn5 gene for kanamycin resistance.The effects of deleting different regions on morphological changes and on patterns of protein synthesis during fruiting body formation were examined. The process of fruiting body formation was severely delayed when both the ops and the tps genes were deleted. However, this delay could be suppressed by either the ops gene or the tps gene, individually, although in the latter case, a slight delay was still observed. These results indicate that the ops gene is expressed during fruiting body formation and plays a role in the normal program of M. xanthus differentiation. Furthermore, the role of the ops gene can be complemented by the tps gene. The deletion of the ops and/or tps genes had no effect on glycerol-spore formation.     

Molecular Cloning and Nucleotide Sequence of the Porphobilinogen Deaminase Gene,hemC, from Chlorobium vibrioforme

We previously reported the DNA sequence and expression of the Chlorobium vibrioforme glutamyl-tRNA reductase (hemA) gene (Majumdar et al., Arch Microbiol 156:281, 1991). The sequence downstream of the hemA gene indicated homology to Escherichia coli and Bacillus subtilis porphobilinogen deaminase (hemC) gene. The Chlorobium gene was confirmed to be the porphobilinogen deaminase gene, and complete sequence of the structural gene was obtained. A 2.8-kb DNA fragment containing the 1.3-kb hemA gene of Chlorobium was cloned into a hemC auxotroph (Sz16) of Bacillus subtilis, and complementation of the auxotroph to prototrophy was achieved. DNA sequence data showed a single open reading frame of 840 bp coding a protein of 279 amino acid residues. The deduced amino acid sequence of the Chlorobium porphobilinogen deaminase revealed 39% to 46% homology with the corresponding prokaryotic and eukaryotic sequences. Received: 20 September 1996 / Accepted: 26 October 1996     

A Method of Determination of Penta-, Tetra-, Tri-, and Disaccharides from Xylan by Ion-exchange Chromatography

Bacillus subtilis B7, a tmrA mutant, shows both tunicamycin resistance and a-amylase hyperproductivity. The tmrA characters can be transferred simultaneously to recipient cells by DNA-mediated transformation. We found a typical gene amplification phenomenon in the tmrA transformants and B7 strain. The amplified unit, 16.3kb in size, covers a-amylase structural gene amyE to another tunicamycin resistance gene tmrB, which is located 9kb downstream of the amyE gene. About 10 repeating units are supposed to be tandemly repeated in the transformants. Amplification of the wild amyE and tmrB genes could be the cause of the α-amylase hyperproductivity and tunicamycin resistance of the tmrA transformants and B7 strain.     

Is the egg size determining gene,Esd, on the W chromosome identical with the sex-linked giant egg gene,Ge, in the silkworm?

The presence of the egg size-determining (Esd) gene, which acts as a quantitative gene, on the W chromosome of the silkworm was revealed in a previous study by using two types of triploid females, ZZW and ZWW, (Kawamura, Genetica 76: 195–201). The females with the sex-linked giant egg (Ge) gene deposit eggs as large as those laid by tetraploids. If the Ge mutant is induced by translocation of a fragment of the W carrying Esd onto the Z by chance, the egg size increase in the Ge strain and in tetraploids may be easily explained by the double dose of Esd. The measurement of the length of the Z-W bivalent in oocytes showed that the Z of the Ge strain was much longer than that of the other strains which do not carry the Ge gene. The result suggests that the Ge gene is identical with the Esd on the W chromosome of the silkworm.     

木薯UGT41基因对细菌性枯萎病的抗性研究北大核心CSCD

糖基转移酶广泛存在于植物中,其中UDP依赖型糖基转移酶(UDP-glycosyltransferases,UGTs)基因家族是糖基转移酶中的一大类。该研究以华南124木薯品种(Manihot esculenta cv.SC124)为材料,利用RT-PCR技术克隆木薯MeUGT41基因,以病毒诱导干扰木薯MeUGT41基因的表达量,并对基因干扰植株进行细菌性枯萎病抗性评价,为研究MeUGT41基因在木薯抵抗细菌性枯萎病的抗病机理奠定基础。结果表明:(1)地毯草黄单胞菌(Xamthomonas axonopodis pv.Manihotis,Xam)可显著诱导木薯MeUGT41基因表达。(2)成功构建MeUGT41的病毒诱导基因沉默(VIGS)载体,将干扰载体转化至木薯叶片进行MeUGT41基因沉默,荧光定量PCR检测结果显示,木薯叶片中MeUGT41基因的表达量显著下降。(3)Xam侵染实验表明,干扰抑制MeUGT41基因表达可显著降低木薯植株叶片对Xam病菌侵染的抵抗能力。研究认为,木薯叶片中MeUGT41基因具有抵抗Xam病菌侵染的能力。     

Isolation,expression, and evolution of the gene encoding mitochondrial elongation factor Tu in Arabidopsis thaliana

We have characterized a second nuclear gene (tufM) in Arabidopsis thaliana that encodes a eubacterial-like protein synthesis elongation factor Tu (EF-Tu). This gene does not closely resemble the previously described Arabidopsis nuclear tufA gene, which encodes the plastid EF-Tu, and does not contain sequence elements found in all cyanobacterial and plastid tufA genes. However, the predicted amino acid sequence includes an N-terminal extension which resembles an organellar targeting sequence and shares three unique sequence elements with mitochondrial EF-Tu's, from Saccharomyces cerevisiae and Homo sapiens, suggesting that this gene encodes the Arabidopsis mitochondrial EF-Tu. Consistent with this interpretation, the gene is expressed at a higher level in flowers than in leaves. Phylogenetic analysis confirms the mitochondrial character of the sequence and indicates that the human, yeast, and Arabidopsis tufM genes have undergone considerably more sequence divergence than their cytoplasmic counterparts, perhaps reflecting a cross-compartmental acceleration of gene evolution for components of the mitochondrial translation apparatus. As previously observed for tufA, the tufM gene is present in one copy in Arabidopsis but in several copies in other species of crucifers.     

Distribution of Pathogenic Genes aatA, aap, aggR, among Uropathogenic Escherichia coli (UPEC) and Their Linkage with StbA Gene

Urinary tract infection (UTI) with E. coli (UPEC) is one of the most common bacterial infections among human beings. In addition to the host predisposing factors, genes are also proposed to have an important role in the occurrence of UTIs. This study investigated the distribution of three pathogenic genes including aggR, aap and aatA among UPEC infected samples and their linkage with stbA, the essential gene for maintaining of pAA plasmid. A total of 244 samples were collected from patients with UTIs through clinical laboratories located in western side of Tehran (Iran) during years 2008–2009. E. coli isolation was performed according to standard laboratory methods. DNAs were extracted from samples using Boiling method, and the presence of aap, aggR, aatA and stbA genes were investigated by PCR. No pathogenic genes (aap, aggR, aatA) were found in 104 out of 244 UPEC samples, while 14 of them were carrying stbA gene. Out of 140 UPEC samples with pathogenic genes, 94 (46.6%) were carrying aap gene, 52 (23%) aggR gene, and 80 (35.4%) aatA gene. A total of 18 samples were also carrying all pathogenic genes together. Moreover, 44 out of 144 samples were carrying stbA gene. The results obtained by this study showed that the aggR, aap and aatA pathogenic genes have different existence patterns in different E. coli strains that infect different organs. Our study also showed that these three plasmid genes in EAEC strains are able to transpose in the genome and change their level of linkage with pAA plasmid essential gene stbA. Meanwhile, this study confirmed that aggR, aap and aatA genes are not specific to only EAEC strains.     

Molecular characterization of intact, but cryptic, flagellin genes in the genus Shigella

Flagellin genes (fliC) were detected in two species of the genus Shigella. The fliC_SF gene cloned from Shigella flexneri produced normal-type flagella in an Escherichia coli δ fliC strain while the fliC_SS genes from two Shigella sonnei strains produced curly-type flagella and their expression is repressible by Salmonella FljA repressor. The fliC_SF gene (1650bp) shared high similarity with the E. coli fliC_E gene not only in the 5′ and 3′ constant sequences but also in the upstream and downstream sequences. The fliC_S genes (1572 bp) shared high similarity with the Salmonella typhimurium fliCs gene in the operator and 3’constant sequences and also shared high similarity with the fliC_E gene in the downstream sequence, suggesting that the fliC gene has undergone horizontal transfer and recombination. Differences In nucleotide sequences of the central variable regions among the four fliC genes, including fliC_E and fliC_S, suggest that they started differentiation in each lineage approximately 80 million years ago. Loss of motility in Shigella seems to be evolutionarily a recent event.     

Identification, cloning, nucleotide sequence and chromosomal map location of hns, the structural gene for Escherichia coli DNA-binding protein H-NS

Summary Beginning with a synthetic oligonucleotide probe derived from its amino acid sequence, we have identified, cloned and sequenced the hns gene encoding H-NS, an abundant Escherichia coli 15 kDa DNA-binding protein with a possible histone-like function. The amino acid sequence of the protein deduced from the nucleotide sequence is in full agreement with that determined for H-NS. By comparison of the restriction map of the cloned gene and of its neighboring regions with the physical map of E. coli K12 as well as by hybridization of the hns gene with restriction fragments derived from the total chromosome, we have located the hns gene oriented counterclockwise at 6.1 min on the E. coli chromosome, just before an IS30 insertion element.     

Cloning, sequencing and expression of a recA gene from Amycolatopsis mediterranei

The 3.5 kb nucleotide fragment, including the recA gene and its downstream recX-like gene, has been isolated from a genomic library by dot-blot hybridization with the Mycobacterium smegmatis recA gene. The recA gene, consisting of 1047 base pairs (bp), encodes a polypeptide of 348 amino acids while the recX-like gene, consisting of 450 bp, encodes a shorter polypeptide of 149 amino acids. Both the deduced amino acid sequences of recA and recX resemble those of the recA and recX genes from other bacteria. The cloned Amycolatopsis mediterranei U32 recA gene conferred partial resistance to ethyl methane sulfonate when expressed in E. coli with the lacZ promoter.     

Chromosomal and regional localization of the loci for IGKC,IGGC, ALDB,HOXB, GPT,and PRNP in the American mink (Mustela vison): comparisons with human and mouse

Chromosomal localization of the genes for gamma- and kappa-immunoglobulins (IGGC and IGKC, respectively), aldolase B (ALDB), prion protein (PRNP), homeo box B (HOXB), and glutamate pyruvate transaminase (GPT) were determined with the use of mink-rodent hybrid cells. Analysis of segregation of the mink markers and chromosomes in these hybrid cells allowed us to assign the gene for HOXE to Chromosome (Chr) 8, IGGC to Chr 10, PRNP and IGKC to Chr 11, ALDB to Chr 12, and GPT to Chr 14 in mink. Furthermore, using a set of mink-mouse hybrid cells carrying fragments of mink Chr 8 of different sizes, we assigned the gene for HOXB to the pter-p26 region of the short arm of Chr 8. Comparative mapping of the genes of mink, human, and mouse, as well as other mammalian species, demonstrated that the mink genes HOXB, PRNP, ALDB, and IGGC are members of a conserved region shared by many mammalian species in common; the IGKC gene is a member of a conserved region common to carnivores and primates, not rodents; the GPT gene is a member of a syntenic gene group probably unique to the Mustelidae family or carnivores.     

Complexity,polymorphism, and connectivity of mouse V k gene families

To define the polymorphism and extent of the mouse immunoglobulin kappa (Igk) gene complex, we have analyzed restriction-enzyme digested genomic DNA from 33 inbred strains of mice with labeled DNA probes corresponding to 16 V _x protein groups (1 of them previously undescribed) and the J _k/C _K region (V, variable; J, joining; C, constant). These probes detected between 1 and 25 distinct restriction enzyme fragments (REF) that appeared in up to eight polymorphic patterns, thus defining eight mouse Jgk haplotypes. The investigated portion of the V _A repertoire was estimated to encompass between 60 and 120 discernable V _k gene-containing REFs. In contrast to mouse V _H gene families, several V _k gene families defined by these probes appeared to overlap. This observation has implications for V _k gene analyses by nucleic acid hybridization and raises the possibility that the V _A gene complex is a continuum of related sequences.Abbreviations used in this paper C constant - Ig immunoglobulin - J joining - REF restriction enzyme fragment - RFLP restriction fragment length polymorphism - V variable     

Isolation,identification, and characterization of <Emphasis Type="Italic">Vibrio cholerae</Emphasis> from the Danube River in Slovakia

The occurrence of Vibrio cholerae, an important aquatic pathogen, was assessed in the surface water of the Danube River near Bratislava. The isolates were distinguished by biochemical tests and grouped by ARDRA to three clusters corresponding to three species (V. cholerae, Vibrio metschnikovii, and Aeromonas spp.). The identification of V. cholerae was confirmed by multiplex PCR using primer pairs targeted to ompW gene (membrane protein), ctxA gene (toxicity gene), and toxR gene (regulatory gene). None from the isolated V. cholerae from surface water contained ctxA gene; seven of them possessed toxR gene. Serotyping of V. cholerae isolates with polyvalent O antiserum and O/139 antiserum was negative. All isolates of V. cholerae were susceptible to chloramphenicol, rifampicin, tetracycline, variable to ampicillin, and resistant to kanamycin and streptomycin.     

Expression of carp-cdx1, a caudal homolog,in embryos of the carp,Cyprinus carpio

A carp caudal cDNA of 1.3 kb was cloned after screening an early segmentation stage cDNA library with a probe produced by PCR using conserved homeobox sequences as primers and genomic DNA as template. The homeobox gene was called carp-cdxl. The gene appears highly similar to other vertebrate caudal homologs, especially the zebrafish gene cdx(Zf-cad). The possible relationship to homeobox genes within the Hox-C gene complexes is discussed. A weak expression of the gene, detected by in situ hybridization, was found shortly before gastrulation (at 25% epiboly) in cells likely to have a posterior fate. During gastrulation expression became stronger. At the early segmentation stage, cells of the neural keel in the area of the prospective spinal cord expressed the gene. During the progression of segmentation, expression retracted in a caudal direction. The tailbud expressed the gene throughout, but the somites lost expression shortly after their formation. Only the most lateral mesoderm cells maintained expression in the trunk area. Carp-cdxl was also expressed in the endoderm. At 24 h after fertilization the gene was only expressed in the tailbud. At 48 h, no expression could be detected. The expression pattern suggests a function for carp-cdxl in gastrulation and patterning along the anterior-posterior axis of the embryo.     

Identification, characterization, and chromosomal organization of the ftsZ gene from Brevibacterium lactofermentum

The ftsZ gene was cloned from the chromosomal DNA of Brevibacterium lactofermentum by the polymerase chain reaction (PCR) using two oligonucleotides designed from two conserved regions found in most of the previously cloned and sequenced ftsZ genes from other microorganisms. ftsZ is a single-copy gene in corynebacteria and is located downstream from ftsQ and murC, indicating linkage between genes involved in peptidoglycan synthesis (mur genes) and genes involved in cell division (fts genes). The organisation of the cluster is similar to that in Streptomyces and different from those of Escherichia coli or Bacillus subtilis because ftsA is not located upstream of ftsZ. The gene was expressed in E. coli using the T7 expression system; the calculated molecular weight of the expressed protein was 50 kDa. Expression of the B. lactofermentum ftsZ gene in E. coli inhibited cell division and led to filamentation. The ftsZ gene of this organism does not complement ftsZ mutations or deletions in E. coli, when cloned on low or high-copy-number vectors. Received: 14 January 1998 / Accepted: 31 March 1998     

Cloning, Sequencing, and Characterization of the Azospirillum brasilense fhuE Gene

The fhuE gene of Escherichia coli encodes the FhuE protein, which is a receptor protein in the coprogen-mediated siderophore iron-transport system. A fhuE gene homologue from Azospirillum brasilense, a nitrogen-fixing soil bacterium that lives in association with the roots of cereal grasses, was cloned, sequenced, and characterized. The A. brasilense fhuE encodes a protein of 802 amino acids with a predicted molecular weight of approximately 87 kDa. The deduced amino-acid sequence showed a high level of homology to the sequences of all the known fhuE gene products. The fhuE mutant was sensitive to iron starvation and defective in coprogen-mediated iron uptake. The mutant failed to express one membrane protein of approximately 78 kDa that was induced by iron starvation in the wild type. Complementation studies showed that the A. brasilense fhuE gene, when present on a low-copy number plasmid, could restore the functions of the mutant. Mutation in fhuE gene did not affect nitrogen fixation.