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[Plant genetic engineering in Monsanto company: from the first laboratory experiments to worldwide practical use].

The history of modern biotechnology of agricultural plants is briefly considered in the article. Methods of genetic transformation and regeneration of transgenic plants as well as the mechanisms of resistance of genetically modified plants to herbicides and pests are discussed. By the example of genetically modified varieties and hybrids there are shown the ways of solving the problem of weeds and pests. The questions of biosafety legislation in different countries are considered.

Biotechnology↗
PubMed2005

Kinetic evaluation of photosensitivity in genetically engineered neurons expressing green algae light-gated channels.

Neurons become photosensitive by genetically introducing one of green algae-derived protein, channelrhodopsin-2 (ChR2). Here, we quantitatively investigated the rapidness of the light-gated current of ChR2 expressed in PC12 cells using blue light-emitting diode (LED) light. The light-gated current consists of two components, inactivating and non-inactivating. The magnitude of inactivating component was almost linearly related to the light intensity. The non-inactivating component showed a tendency to saturate at high illumination. Both the activation and inactivation rates of the light-gated current were linearly dependent on the light intensity. However, the activation rate (turning-on rate) is about 10-fold faster than the inactivation rate. Although the turning-off time constant was little dependent on the light intensity, that at the end of 1s light pulse was about two-fold larger than that at 20 ms. Neurons are also made photosensitive by the expression of ChR2 in the living animal. Since both the turning-on and turning-off time constants of light-gated current was smaller than the membrane time constant of neurons, the LED light illumination of the photosensitive neurons was enough to evoke action potentials in a pulse-to-pulse manner in an acute slice of hippocampus.

Algorithms↗
PubMed1987

Implantation of genetically engineered fibroblasts into mice: implications for gene therapy.

In a variety of human genetic diseases, replacement of the absent or defective protein provides significant therapeutic benefits. As a model for a somatic cell gene therapy system, cultured murine fibroblasts were transfected with a human growth hormone (hGH) fusion gene and cells from one of the resulting clonal lines were subsequently implanted into various locations in mice. Such implants synthesized and secreted hGH, which was detectable in the serum. The function of the implants depended on their location and size, and on the histocompatibility of the donor cells with their recipients. The expression of hGH could be modified by addition of regulatory effectors, and, with appropriate immunosuppression, the implants survived for more than 3 months. This approach to gene therapy, here termed "transkaryotic implantation," is potentially applicable to many genetic diseases in that the transfected cell line can be extensively characterized prior to implantation, several anatomical sites are suitable for implantation, and regulated expression of the gene of therapeutic interest can be obtained.

Animals↗
PubMed1981

Site-specific insertion of genes into T-DNA of the Agrobacterium tumor-inducing plasmid: an approach to genetic engineering of higher plant cells.

This paper presents a method for insertion of genetic material into a specific site in T-DNA, the portion of Agrobacterium tumor-inducing (Ti) plasmid that becomes incorporated into the nuclear DNA of transformed plant cells when crown gall tumors are incited by this plant pathogen. The three stages of our procedure are as follows: 1. A T-DNA subfragment cloned in pBR322 is cleaved by a restriction endonuclease at a unique central site and target DNA (a kanamycin resistance marker) is ligated into this site. 2. The resulting recombinant plasmid is purified and cleaved with EcoRI; the resulting fragment bearing the kanamycin resistance marker is ligated into the unique EcoRI site of pRK290, a wide-host-range plasmid. 3. The pRK290 recombinant plasmid is transformed into an agrobacterium tumefaciens strain containing a wild type Ti plasmid. Double recombination between the altered T-DNA fragment of the clone and its wild-type counterpart in the Ti plasmid is selected for by introduction of R751-pMG2, a plasmid incompatible with pRK290. The approach described here can be adapted for introducing genes into higher plant cells with the Ti plasmid as vector. It can likewise be used for site- or fragment-specific mutagenesis of the Ti plasmid as a means of detailed functional analysis.

DNA↗