RESUMO
Several regulatory organisations are involved in the assessment of clinical gene therapy trials involving genetically modified organisms (GMOs) in The Netherlands. Medical, ethical and scientific aspects are, for instance, evaluated by the Central Committee on Research Involving Human Subjects (CCMO). The Ministry of Housing, Spatial Planning and the Environment (VROM) is the competent authority for the environmental risk assessment according to the deliberate release Directive 2001/18/EC. A Gene Therapy Office has been established in order to streamline the different national review processes and to enable the official procedures to be completed as quickly as possible. Although the Gene Therapy Office improved the application process at the national level, there is a difference of opinion between the EU member states with respect to the EU Directive according to which gene therapy trials are assessed, that urges for harmonisation. This review summarises the gene therapy legislation in The Netherlands and in particular The Netherlands rationale to follow Directive 2001/18/EC for the environmental risk assessment.
Assuntos
Terapia Genética/legislação & jurisprudência , Ensaios Clínicos como Assunto , Comitês de Ética em Pesquisa , União Europeia , Regulamentação Governamental , Humanos , Países Baixos , Organismos Geneticamente Modificados , Medição de RiscoRESUMO
Transformation of uropathogenic Escherichia coli strains with plasmid DNA was in general unsuccessful or very inefficient. Transformation was much more efficient when galE mutants of such strains, in which the lipopolysaccharide chains appeared shorter, were used as recipients.
Assuntos
Escherichia coli/genética , Genes Bacterianos , Lipopolissacarídeos/fisiologia , Plasmídeos , Transformação Bacteriana , Escherichia coli/patogenicidade , Galactose/metabolismo , Lipopolissacarídeos/análise , Mutação , Sistema Urinário/microbiologia , VirulênciaRESUMO
Cloning techniques make it possible to accommodate bacterial genes on vector DNA molecules. On that basis the investigation of bacterial structures and functions got new impetus. The potentials of molecular genetics for detailed analysis of bacterial structures are illustrated in this paper for the gene cluster involved in the expression of F72 fimbriae associated with a uropathogenic Escherichia coli O6:K2:H1:F7 strain.
Assuntos
Escherichia coli/genética , Fímbrias Bacterianas , Genes Bacterianos , Proteínas de Bactérias/genética , Sequência de Bases , Clonagem Molecular , DNA Bacteriano/análise , DNA Recombinante/análise , Hibridização de Ácido Nucleico , NucleotídeosRESUMO
By cloning a distinct 8 MDa fragment of the IncI plasmid, R144, in the vector pACYC184, two recombinant plasmids were isolated. In these plasmids, pRAH303 and pRAH308, the inserted fragment was in opposite orientations. Both plasmids when present in a recipient strain caused a conjugation-specific exclusion in crosses with donor cells carrying the IncI plasmid R144. Some derivatives of the recombinant plasmids in which parts were deleted, or in which Tn5 transposons were inserted, appeared to be exclusion negative. Analysis in minicells of the gene products of such plasmids together with those of the original recombinant plasmids revealed that the presence of two proteins, with apparent molecular weights of 13,000 and 19,000 Da could be correlated with the exclusion phenomenon.
Assuntos
Conjugação Genética , Plasmídeos , Proteínas de Bactérias/genética , Clonagem Molecular , Elementos de DNA Transponíveis , DNA Bacteriano/genética , Escherichia coli/genética , Genes Bacterianos , Peso MolecularRESUMO
In Escherichia coli transformation with linear donor deoxyribonucleic acid, the recBC pathway is functional, but genetic analysis shows that the recBC nuclease is deleterious to linear deoxyribonucleic acid.
Assuntos
Desoxirribonucleases/fisiologia , Proteínas de Escherichia coli , Escherichia coli/genética , Transformação Bacteriana , DNA Bacteriano/genética , Exodesoxirribonuclease V , Recombinação GenéticaRESUMO
E. coli K12 transformants, selected as leu+ or pyrA+ transformants, were analysed for inheritance of some closely linked unselected markers. Based on the observation that the number of recombinants which require, besides an integration event, one or more crossing-over events was negligible, a simple mapping function was deduced. The function L= e-kd, which directly relates observed linkage of an unselected marker and the relative distance of that unselected marker to the selected marker, gave a consistent interpretation of the experimental results.
