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  • 1
    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Translational fusions between a mutant phoA (lacking its promoter, ribosomal binding site and signal peptide sequence) and Rhizobium‘symbiotic’ genes were isolated. Since these fusions expressed alkaline phosphatase (AP), the product of phoA, the genes into which phoA was inserted apparently specify proteins located in the bacterial periplasm or cell membrane, the compartment in which AP has activity. These genes were psiA and genes upstream of psiA (psiA is required for normal nodule development and strains with multicopy psiA fall to make exopolysaccharide (EPS) and to nodulate). Fusions between phoA and pss (exo) genes, which are required for EPS production, also resulted in the expression of AP indicating that products of these pss genes were located at the cell surface. Using grus fusions to psiA and pssA, we found that the former was expressed in N2-fixing bean root nodules but the latter was not.
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  • 2
    ISSN: 1573-5028
    Keywords: oxygen-evolving complex ; woxA, Anabaena ; manganese-stabilizing polypeptide ; photosystem II
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The 33 kDa extrinsic polypeptide of photosystem II, also known as the manganese-stabilizing polypeptide (MSP), is located on the lumen side of the thylakoid and is involved in water oxidation. The gene for MSP, designated woxA, has been cloned from the nitrogen-fixing filamentous cyanobacterium Anabaena and sequenced. The woxA open reading frame was found to be 819 bp. The deduced amino acid sequence was 63% and 59% homologous with that of Synechococcus and Synechocystis, respectively, and 44% conserved when compared to the MSP of spinach or pea. Two cysteine residues at positions 48 and 73 were found to be conserved in cyanobacteria and plants. The first 29 amino acids are hydrophobic and may represent the transit peptide. woxA::phoA translational fusion products, in which the body of Escherichia coli alkaline phosphatase was fused to the amino terminal portion of woxA between residues 35 and 130, yielded active alkaline phosphatase in E. coli. Thus the transit peptide of woxA functions in E. coli to transport phosphatase across the cytoplasmic membrane. S1 mapping and primer extension experiments showed that the woxA transcription intitiation site is located 220 bp upstream from the translational start. The woxA promoter has some resemblance to the E. coli consensus and other known Anabaena vegetative cell promoters.
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    Theoretical and applied genetics 74 (1987), S. 95-99 
    ISSN: 1432-2242
    Keywords: Microspore ; Anther culture ; Haploid production ; Oryza sativa L. ssp. indica
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary High frequencies of callusing were obtained in indica rice from the microspores which were staged in acetic acid iron alum-haematoxylin stain prior to culture on G5 medium. Two local varieties, ‘Khonorullo’ and ‘Namyi’, and two advance pre-release cultures, ‘PK 1-1-3’ and ‘PK 12-22’, were used in this investigation. All the cultures exhibited a wide adaptation to varying medium; however, the frequency of callusing was highest (45.5%) in ‘PK 1-1-3’ followed by ‘PK 12-22’ (32.4%) and ‘Khonorullo’ (31.6%). Cold shock (10 °C) for 11 days enhanced the frequency of callusing by 200% in ‘Khonorullo’.
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  • 4
    ISSN: 1617-4623
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A strain of R. phaseoli cured of its symbiotic plasmid, pRP2JI, retained the ability to make exopolysaccharide (EPS). However, a region of pRP2JI, when cloned at an increased copy number in wide host-range vectors and transferred to this and other strains of Rhizobium, inhibited EPS synthesis. The gene responsible was termed psi (polysaccharide inhibition) and was located in a region of the symbiotic plasmid close to nodulation and nitrogen fixation genes. psi is important in the symbiosis since a wild-type strain containing psi cloned on a multicopy plasmid failed to form Phaseolus nodules, and mutant strains containing psi::Tn5 mutations failed to fix nitrogen in Phaseolus nodules. It is proposed that the function of psi may be to repress in the bacteriod the expression of genes such as those for EPS synthesis which are normally expressed in free-living culture.
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  • 5
    ISSN: 1617-4623
    Keywords: Gene regulation ; Melanin synthesis ; Nitrogen fixation ; Phaseolus beans ; Rhizobium phaseoli
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The symbiotic plasmid pRP2JI of Rhizobium phaseoli strain 8002 was shown to contain two separate regions of DNA which are required and sufficient for the synthesis of the pigment melanin. One of these regions containing the class II mel gene(s) was located to other genes involved in nodulation and in nitrogen fixation. Mutations in this region abolished both the ability to synthesize melanin and to fix nitrogen in Phaseolus bean root nodules. Mutations in the other, unlinked region, containing class I mel gene(s), also abolished melanin synthesis but did not affect symbiotic nitrogen fixation. Transcriptional fusions between the class I mel gene and the Escherichia coli lacZ gene were constructed and it was demonstrated that the class II mel gene(s) activated their transcription in free-living culture. Further, strains containing the cloned regulatory class II gene(s) synthesized melanin when growing in minimal medium, in contrast to wild-type strains which became pigmented only in complete medium containing yeast extract and tryptone. It was shown by hybridization experiments that the regulatory mel gene was closely linked to or may correspond to the regulatory nifA gene; a fragment of R. phaseoli DNA which included the class II gene(s) of R. phaseoli hybridized to a previously identified nifA-like gene of R. leguminosarum, the species that nodulates peas.
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  • 6
    ISSN: 1617-4623
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
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  • 7
    ISSN: 1617-4623
    Keywords: DNA sequence ; Exopolysaccharide ; Nodulation ; psi ; pss ; Rhizobium leguminosarum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Strains of Rhizobium leguminosarum (R. l.) biovar viciae containing pss mutations fail to make the acidic exopolysaccharides (EPS) and are unable to nodulate peas. It was found that they also failed to nodulate Vicia hirsuta, another host of this biovar. When peas were co-inoculated with pss mutant derivatives of a strain of R.l. bv viciae containing a sym plasmid plus a cured strain lacking a sym plasmid (and which is thus Nod-, but for different reasons) but which makes the acidic EPS, normal numbers of nodules were formed, the majority of which failed to fix nitrogen (the occasional Fix+ nodules were pressumably induced by strains that arose as a result of genetic exchange between cells of the two inoculants in the rhizosphere). Bacteria from the Fix- nodules contained, exclusively, the strain lacking its sym plasmid. When pss mutant strains were co-inoculated with a Nod- strain with a mutation in the regulatory gene nodD (which is on the sym plasmid pRL1JI), normal numbers of Fix+ nodules were formed, all of which were occupiced solely by the nodD mutant strain. Since a mutation in nodD abolishes activation of other nod genes required for early stages of infection, these nod genes appear to be dispensable for subsequent stages in nodule development. Recombinant plasmids, containing cloned pss genes, overcame the inhibitory effects of psi, a gene which when cloned in the plasmid vector pKT230, inhibits both EPS production and nodulation ability. Determination of the sequence of the pss DNA showed that one, or perhaps two, genes are required for correcting strains that either carry pss mutations or contain multi-copy psi. The predicted polypeptide product of one of the pss genes had a hydrophobic aminoterminal region, suggesting that it may be located in the membrane. Since the psi gene product may also be associated with the bacterial membrane, the products of psi and pss may interact with each other.
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  • 8
    Electronic Resource
    Electronic Resource
    Springer
    Journal of applied phycology 7 (1995), S. 561-564 
    ISSN: 1573-5176
    Keywords: Spirulina ; cyanobacteria ; restriction enzyme
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Four unique restriction enzymes were identified in the soluble protein fraction of Spirulina platensis strain pacifica, a commercially important strain of marine cyanobacterium that is used as a supplement in a human diets. These are SpaI, SpaII, SpaIII and SpaIV, which are isoschizomers of Tth111I, Pvul, PvuII and HindIII, respectively. The recognition sites of each of these four enzymes were identified by restriction digests of different plasmid DNAs of known sequence and determining the cleavage sites by sequencing. SpaI is the most predominant restriction enzyme present in S. platensis strain pacifica. It shows high activity at 37 °C compared to 65 °C for its isoschizomer Tth111I.
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  • 9
    Electronic Resource
    Electronic Resource
    Springer
    Journal of applied phycology 8 (1996), S. 259-259 
    ISSN: 1573-5176
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
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  • 10
    ISSN: 1617-4623
    Keywords: Exopolysaccharide ; Legume ; Nodules ; Rhizobium ; Xanthomonas
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A Tn5-induced mutant strain of R. phaseoli which failed to synthesize exopolysaccharide (EPS) was isolated and was shown to induce normal nitrogen-fixing nodules on Phaseolus beans, the host of this Rhizobium species. The corresponding wild-type Rhizobium DNA was cloned in a wide host-range vector and by isolating Tn5 insertions in this cloned DNA, mutations in a gene termed pss (polysaccharide synthesis) were isolated. These were introduced by marker exchange into near-isogenic strains of R. leguminosarum and R. phaseoli which differed only in the identity of their symbiotic plasmids. Whereas the EPS-deficient mutant strain of R. phaseoli induced normal nitrogen-fixing nodules on Phaseolus beans, the same mutation prevented nodulation of peas by a strain of R. leguminosarum which normally nodulates this host. Further, it was found that DNA cloned from the plant pathogen Xanthomonas campestris pathover campestris could correct the defect in EPS synthesis in R. leguminosarum and R. phaseoli and also restored the ability to nodulate peas to the pss::Tn5 mutant strain of R. leguminosarum.
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