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Instituto de Biologia Unicamp







ABSTRACTA (3) 2000

Artigos Publicados em Periódicos

P100-00 Expression of correctly processed human growth hormone in seeds of transgenic tobacco plants Leite A, Kemper EL, Silva MJ, Luchessi AD, Siloto RMP, Bonaccorsi ED, El-Dorry H, Arruda P* Human growth hormone was expressed in transgenic tobacco seeds using the monocot tissue-specific promoter from sorghum gamma-kafirin seed storage protein gene. During tobacco seed ripening, the expressed hormone was directed to the endoplasmic reticulum by a signal peptide from a Coix prolamin and was secreted into the apoplastic space, where it accounted for 0.16% of total soluble seed protein. The expressed hormone has the same amino acid sequence and receptor-binding properties as the native mature hormone. Molecular Breeding 6: 47-53, 2000. IF = 2.786 *E-mail:

P101-00 Making colony PCR easier by adding gel-loading buffer to the amplification reaction Menossi M*, Cremonese N, Maron LG, Arruda P* Here we evaluated the effects of three widely used gel-loading buffers on the amplification of targets with different sizes, from different sources and using several primer combinations. Biotechniques 28: 424-425, 2000. IF = 1.749 *E-mail:;
P102-00 Molecular evidence that sugarcane yellow leaf virus (ScYLV) is a member of the Luteoviridae family Maia IG, Gonçalves MC, Arruda P*, Vega J* A previously uncharacterized virus was reported in southeast Brazil causing a yellowing leaf disease in sugarcane. The virus, termed sugarcane yellow leaf virus (ScYLV), shares features typical of the luteoviruses. The nucleotide sequence of the coat protein (CP), 17 kDa protein and C-tenminus of the RNA-dependent RNA polymerase coding regions was determined from an RT-PCR amplification product. Comparisons showed that the deduced amino acid sequences share a considerable degree of identity and similarity with corresponding sequences of known luteoviruses, thus clearly establishing ScYLV as a member of the family Luteoviridae. The authenticity of the CP open reading frame was confirmed by its expression in E. coli. The recombinant CP positively reacted in immunoblot assays with polyclonal antibodies raised against native ScYLV. Phylogenetic analyses also suggest that the 5' and 3' coding blocks of the ScYLV genome possess different taxonomic affinities within the Luteoviridae family, as does also the genome of soybean dwarf virus. Archives of Virology 145: 1009-1019, 2000. IF = 1.591 *E-mail:;
P103-00 Regulation of lysine catabolism in higher plants Arruda P*, Kemper EL, Papes F and Leite A Lysine is an important essential amino acid for mammals, but its concentration in cereals, one our main food sources, is low. Research over the past 40 years has unraveled many biochemical and molecular details of the aspartic acid pathway, which is the main route for lysine biosynthesis in plants. However, genetic manipulation of this pathway has not been successful at producing high-lysine seeds. This is because lysine, instead of being accumulated, is degraded via the saccharopine pathway. Recent work has increased our knowledge of this pathway, including both the enzymes involved and their regulation. Trends in Plant Science 5: 324-330, 2000. IF = 9.350 *E-mail:
P104-00 Degradation of lysine in rice seeds: Effect of calcium, ionic strength, S-adenosylmethionine and S-2-aminoethyl-L-cysteine on the lysine 2-oxoglutarate reductase-saccharopine dehydrogenase bifunctional enzyme Gaziola SA, Sodek L, Arruda P*, Lea PJ and Azevedo RA Lysine biosynthesis has been extensively studied and the regulatory enzymes characterized in some of the most important crop plants, however, much less is known about the lysine degradation pathway. Lysine 2-oxoglutarate reductase (LOR) and saccharopine dehydrogenase (SDN) have recently been partially purified and characterized from plants, and have been shown to exist as a single bifunctional polypeptide, We have further characterized these enzymes from rice endosperm in relation to Ca2+ and ionic strength modulation, Optimum pH values of 7.0 and 8.0 were obtained for LOR and SDH, respectively, It would appear that the modulation by Ca2+ and ionic strength of LOR is a common feature among plant LOR enzymes, S-adenosylmethionine did not produce any significant effect on either enzyme activity, indicating that it only plays a role in the regulation of lysine biosynthesis. The effect of S-2-aminoethyl-L-cysteine (AEC) as both a substrate and an inhibitor of LOR activity was also tested, AEC was shown to partially substitute for lysine as a substrate for LOR, but was also able to inhibit LOR activity. The higher K-m for AEC compared to lysine may reflect a lower binding affinity for AEC. Physiologia Plantarum, 110: 164- 171, 2000. IF = 2.460 *
P105-00 The mitochondrial genome of the primary screwworm fly Cochliomyia hominivorax (Diptera: Calliphoridae) Lessinger AC, Martins Junqueira AC, Lemos TA, Kemper EL, Da Silva FR, Vettore AL, Arruda P, L Azeredo-Espin AM* The complete sequence of the mitochondrial genome of the screwworm Cochliomyia hominivorax was determined. This genome is 16 022 bp in size and corresponds to a typical Brachycera mtDNA. The nucleotide composition of C. hominivorax mtDNA is 77% AT-rich, reflected in the predominance of AT-rich codons in protein-coding genes. Non-optimal codon usage was commonly observed in C. hominivorax mitochondrial genes. Phylogenetic analysis distributed the Acalypterate species as a monophyletic group and assembled the C. hominivorax (Calyptratae) and the Acalyptratae in a typical Brachycera cluster. The identification of diagnostic restriction sites on the sequenced mitochondrial genome and the correlation with previous RFLP analysis are discussed. Insect Molecular Biology 5:521-529, 2000. IF = 2.412 *E-mail:
P106-00 An Arabidopsis gene induced by wounding functionaly homologous to flavoprotein oxidoreductases Costa CL, Arruda P*, Benedetti CE The regulation of genes in response to wounding is mediated in part by the octadecanoids 12-oxo-phytodienoic acid (OPDA), jasmonic acid (JA) and its methyl ester methyl jasmonate (MeJA). We identified, by differential display, an Arabidopsis gene (OPR3) induced after wounding. OPR3 is homologous to members of the flavin mononucleotide (FMN) binding proteins. Transcripts of OPR3 rapidly accumulated in leaves after wounding and MeJA treatment, but they were detected in various tissues of unwounded plants at relatively low levels. Expression of the OPR3 gene was significantly reduced in wounded leaves of the coi1 mutant, indicating partial dependence on jasmonate perception for full induction of the gene. The recombinant OPR3 protein also showed beta -NADPH oxidation activity in the presence of cyclohexenone, but not cyclohexanone, suggesting that the enzyme has specificity to cleavage of olefinic bonds in cyclic enones. The results show that the OPR3 gene product represents a new OPR of Arabidopsis induced after wounding. Plant Molecular Biology, 44: 61-71, 2000. IF = 3.102 *E-mail:
P107-00 Improved analysis of promoter activity in biolistically transformed plant cells Menossi M, Puigdomènech P, Martínez-Izquierdo JA Transient assays using particle bombardment are a powerful way to rapidly evaluate gene expression in intact plant tissues. We developed an approach to decrease data dispersion and also evaluated the consistency of the two assays of GUS activity used to determine the promoter activity in transient assays. This approach uses three genes (C1, B-Peru and GFP) and decreases data variability considerably. Moreover, it avoids a loss of time and money in the evaluation of non transformed and poorly transformed samples. Our results also show that the histochemical spot counting method for the quantification of promoter-driven GUS activity, may understimate the activity of strong promoters. For this reason, the fluorometric assay is to be preferred when quantifying promoter-driven GUS activity. Biotechniques 28: 54-58, 2000. IF = 1.749 *E-mail:
P108-00 Total lipids and fatty acids of strains of Metarhizium anisopliae Pupin AM, Messias CL*, Piedrabuena AE, Roberts DW Two growth stages, conidia (C) and mycelium (M), and two media, minimal medium (MM) and complete medium (MC), were compared in 10 strains of M. anisopliae, and two strains of M. anisopliae var. majus were similar in percentages of total lipids. Tukey test for average of lipid content in conidia (C) and mycelia (M) cultured on minimal (MM) and complete (MC) media showed significant differences between means at the 5% level for mycelia and conidia, indicating variability in total lipid production and storage during growth. Strains 5 and 7, both variety majus, did not present sizable differences from variety anisopliae. For fatty acids, C18:1 and C18:2, oleic and linoleic, respectively, the differences were all highly significant (p= 1%) with the highest means being obtained for conidia for fatty acid C18:1 and for myclelia for fatty acid C18:2. Brazilian Journal of Microbiology 31:121-128, 2000. *E-mail:
P109-00 Violacein cytotoxicity and induction of apoptosis in V79 cells Melo PD, Maria SS, Vidal BC, Haun M*, Duran N Cytotoxicity and induction of apoptosis by violacein, a pigment potentially effective for the treatment of Chagas' disease and leukemia and lymphoma cells in culture were studied in V79 cells subjected to the TUNEL assay and the Feulgen reaction followed by image analysis. The drug was found to trigger apoptosis but not necrosis, strengthening its potential as a therapeutic agent. In Vitro Cellular & Developmental Biology - Animal 36(8): 539-543, 2000. IF = 1.066 *E-mail:

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