RESUMO
We describe the application of the newly developed RHIZOSCAN software for accurate morphological analysis and determination of overall and local secondary metabolite concentrations in two clones of Beta vulgaris throughout the growth period. Local secondary metabolite concentrations may be determined at any point in the root, and pigment gradients in each lateral root can be followed during culture and saved to the computer. Throughout the entire analysis process, an image appears in a graphical result window on the screen, which enables visual evaluation of the numerical output at each stage of the analysis. Biosynthetic data on concentrations obtained by image analysis were validated by spectrophotometric analysis. The importance of determining appropriate scanning and analysis conditions (scanning resolution, background color, threshold value, segmentation plane in the hue-saturation-intensity color system and pruning length) for obtaining accurate morphological measurements is examined and the means of fixing these parameters is described. Our results show that, using RHIZOSCAN, detailed and accurate information on root architecture and secondary metabolite concentrations can be obtained in a short time.
Assuntos
Automação , Chenopodiaceae/genética , Processamento de Imagem Assistida por Computador/métodos , Software , Chenopodiaceae/metabolismo , Pigmentos Biológicos/química , EspectrofotometriaRESUMO
A Gag protein segment of human immunodeficiency virus 1 (HIV-1) has been fused to a C terminally truncated core antigen of hepatitis B virus (HBcAg) using an E. coli expression system. Fusion of 90 amino acids of HIV-1 Gag protein to HBcAg still allowed the formation of capsids presenting on their surface epitopes of HIV-1 core protein, whereas fusion of 317, 189, or 100 amino acids of Gag prevented self-assembly of chimeric particles. Mice immunized with recombinant particles emulsified with Freund's complete adjuvant (CFA) or aluminium hydroxide developed high anti-HBcAg titers. However, anti-HIVp24 antibodies were detected only in mice inoculated with immunogen emulsified with CFA.
Assuntos
Anticorpos Anti-HIV/biossíntese , Proteína do Núcleo p24 do HIV/imunologia , Anticorpos Anti-Hepatite B/biossíntese , Antígenos do Núcleo do Vírus da Hepatite B/imunologia , Proteínas Recombinantes de Fusão/imunologia , Vacinas contra a AIDS/genética , Vacinas contra a AIDS/imunologia , Animais , Western Blotting , Epitopos/genética , Epitopos/imunologia , Escherichia coli/genética , Adjuvante de Freund , Proteína do Núcleo p24 do HIV/genética , HIV-1/genética , HIV-1/imunologia , Antígenos do Núcleo do Vírus da Hepatite B/genética , Vacinas contra Hepatite B , Vírus da Hepatite B/genética , Vírus da Hepatite B/imunologia , Humanos , Camundongos , Microscopia Eletrônica , Plasmídeos/genética , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/ultraestrutura , Vacinas Sintéticas/genética , Vacinas Sintéticas/imunologia , Vacinas contra Hepatite Viral/genética , Vacinas contra Hepatite Viral/imunologiaRESUMO
Insertion of 48 amino acid long sequence of envelope protein gp51 of bovine leukemia virus (BLV), located from position 56 till 103 of mature protein, into Pro144 position of hepatitis B core antigen (HBcAg) leads to the formation of chimeric capsids. These capsids preserve morphology of intact HBcAg but expose on their outer surface BLV epitopes which are localised in the inserted gp51 fragment and responsible for the recognition of chimeras by monoclonal anti-gp51 antibodies MAK14. The anti-genicity of gp51 epitopes within chimeric capsids is not disturbed after shortening of C terminal part of inserted gp51 fragment by deletion of amino acids 73-103. The resulting chimeras show the same capsid-forming ability as well as HBcAg and gp51 antigenic properties.
