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PubMed1988

An extract from Prospects and Risks of Gene Technology: The Report of the Enquete Commission to the Bundestag of the Federal Republic of Germany.

We are pleased to publish an English translation of the crucial section on germ line gene therapy from the Report of the German Enquete Commission. This section of the Report raises basic ethical questions concerning embryo research, as well as more specific questions relating to germ line gene therapy. This form of gene therapy involves genetic modification of the germ cells. Such a modification would be passed on to any descendents of the person whose genes had been altered. The Report had previously dealt with somatic gene therapy, which does not affect the germ line and hence is not passed on to descendents.

Advisory Committees↗
PubMed2002

Evaluation of gene-technological and conventional methods in the identification of Streptococcus pneumoniae.

To find reliable methods able to identify the "difficult" Streptococcus pneumoniae isolates, an in-house polymerase chain reaction (PCR) for pneumolysin gene (Ply-PCR) and a commercial RNA hybridisation test (AccuProbe) were evaluated. Selected isolates of suspected pneumococci, sent for confirmation of identification and for serotyping, were classified into four groups based on their optochin sensitivity and capsule reaction. All isolates in Group 1, which consisted of 24 typical, optochin-sensitive, encapsulated pneumococcal strains, were positive in the Ply-PCR and AccuProbe tests. In Group 2, which consisted of 25 optochin-sensitive, but unencapsulated pneumococcal strains, all the isolates were positive in the Ply-PCR test, and 23 were positive in the AccuProbe test. In Group 3, which consisted of 15 atypical, optochin-resistant but encapsulated pneumococci, 12 of the isolates were positive in the Ply-PCR and 12 in the AccuProbe test, and 11 of these 12 strains were positive in both tests. In Group 4, which consisted of 36 equivocal optochin-resistant, unencapsulated isolates, 15 strains were positive in the Ply-PCR test and 8 strains in the AccuProbe test. As a conclusion, the Ply-PCR and AccuProbe tests identified similarly typical optochin-sensitive pneumococci, but gave partly controversial results about atypical pneumococci. Thus, they did not reliably help in the identification of suspected pneumococcal isolates lacking the conventional characteristics of pneumococcus.

Bacterial Capsules↗
PubMed1980

[Applied gene technology using biological N2-fixation for an example (author's transl)].

About biological nitrogen fixation most information on the genetic and enzymatic level is known from investigations with Klebsiella pneumoniae. The gene region for N2-fixation (nif) was transferred from K.pneumoniae to the enterobacterium Escherichia coli. After acquisition of nif E.coli is able to fix nitrogen. Plasmid pRD1 was constructed which carries the nif-region. Using genetic engineering techniques the exact location of nif on pRD1 was determined. Subsequently the nif-region comprising 7 operons with 14 known genes was cloned on a multicopy plasmid vector. The use of the Klebsiella-nif-region in biotechnology is discussed. Difficulties arising in experiments designed to construct N2-fixing plants are mentioned.

Cloning, Molecular↗
PubMed1989

[Gene technology in the prevention of cancer].

Cancer may develop in connection with rare, genetic diseases or after exceptional, environmental exposures. Most commonly, however, cancer results from an unhappy coincidence of normal, genetic factors and ordinary, environmental conditions. Oncogenes are normal growth genes. Anti-oncogenes are normal regulatory genes which control growth genes. Cancer may result from changes (mutations) in an oncogene (such as in the Philadelphia chromosome), or from the lack of control of normal oncogenes. The latter may be due to loss of function of a suppressor gene (mutation in an anti-oncogene, such as in retinoblastoma) or through a change in the position of an oncogene (such as in Burkitt's lymphoma and in certain leukemias).

Biomarkers, Tumor↗
PubMed1980

[Possible therapy with and limits of gene technology (author's transl)].

This paper is an attempt to discuss hopes and limits of recent advances in genetechnology especially in respect of their medical applications. Although advances in modern biology are enormous no realistic prognoses can be given at the moment to their medical application in correcting inborn errors of metabolism at the level of the gene. However the production by fermentation of biosubstances like insulin, interferon and hormones, just to name a few, is not only possible but already carried out in many laboratories in USA. Therefore therapy of clinical manifestations like metabolic disorders, viral infections and hormone insufficiencies should become feasible based on the nearly unlimited in vitro production of the necessary compounds.

Animals↗
PubMed2005

High-throughput screening using beta-lactamase reporter-gene technology for identification of low-molecular-weight antagonists of the human gonadotropin releasing hormone receptor.

G-protein coupled receptors (GPCRs) signal via G-proteins to intracellular second messengers. Assays that link transcription of a detectable reporter to promoters that are activated by such signaling cascades are highly sensitive and allow screening for compounds that either activate or inactivate a GPCR of interest. This study describes the development and performance of an antagonistic screen on the human gonadotropin releasing hormone receptor (GnRH-R). Compounds (245,000) were tested in a high-throughput screen using a Chinese hamster ovary cell line stably expressing the human GnRH-R and the Ca2+ sensitive reporter nuclear factor activated in T-cells/ activator protein-1-beta-lactamase. In total, 4,160 active compounds were identified. Colored and toxic compounds, as well as dust and compound aggregates, have been depicted as artifacts. To deselect non-target hits, several follow-up assays, including luminescent and fluorescent Ca2+ mobilization assays and radioligand binding, were developed for the GnRH-R. These assays were validated using peptide and low-molecular-weight GnRH-R reference compounds before hits from screening were also profiled in these assays. For several reference compounds the use of different assay technologies resulted in a poor correlation of potency values. In conclusion, beta-lactamase as a primary high-throughput screening assay is a powerful complementation to other screening technologies. The beta-lactamase technology has several advantages, including lack of cell lysis and ratiometric read-out, which augments assay robustness. Based on technology comparison, it is not adequate to assume that the same hits would be found regardless of which assay technology is used.

Animals↗
PubMed1997

[Application of gene technology in the diagnosis of familial hypercholesterolemia].

Familial hypercholesterolaemia is an autosomal dominant disorder characterized by hypercholesterolaemia, xanthomas and premature coronary heart disease. Treatment of hypercholesterolemia is effective and consists of dietary changes and lipid lowering drugs. Only a minor proportion of familial hypercholesterolaemia patients are adequately treated, however. One explanation for this is assumed to be the relatively vague clinical diagnostic criteria applied. Because familial hypercholesterolaemia is caused by a mutation in the gene encoding the low density lipoprotein (LDL) receptor, mutation analysis of this gene could form the basis for specific diagnosis. 29 different mutations in the LDL receptor gene have been found to cause familial hypercholesterolaemia among Norwegian patients, and a total of 681 patients from 322 unrelated families have been provided with a molecular genetic diagnosis. We conclude that the use of molecular genetic analysis is feasible, and should be used clinically.

DNA Mutational Analysis↗
PubMed1993

Enzymology and gene technology of Streptomyces nucleotide 3'-pyrophosphokinase-2',3'-cyclic monophosphokinase (PPKase).

Streptomyces extracellular nucleotide 3'-pyrophosphokinase-2',3'-cyclic monophosphokinase transfers 5'-beta,gamma-pyrophosphate group from ATP, some ATP derivatives and dATP to a variety of nucleot(s)ides at the 3'-OH site and synthesizes the respective 3'-pyrophosphoryl nucleotides. The enzyme also utilizes A-5'-pn (n = 3-5)-5'-N, transferring their adenosine 5'-pyrophosphoryl group and concomitantly eliminating the AMP moiety therefrom, leading to the synthesis of 2',3'-cyclic monophosphoryl nucleotides. The enzyme gene and its neighbouring up- and downstream sequences were analyzed. Streptomyces and enteric bacteria E. coli and Klebsiella pneumoniae transformants were constructed by incorporation of the pyrophosphokinase gene-recombined expression plasmids. Various effects of the treatments--intracellular synthesis of 3'-pyrophosphoryl nucleotides, retarded cellular growth, stimulated N2 fixation by K. pneumoniae and so on--were seen.

Adenine Nucleotides↗