Abstract: An improved double Recoverable Block of Function (RBF) system for transgene containment with enhanced reliability is disclosed. The system includes two blocking constructs (BCs) at the opposite sides of a transgene of interest (TGI). The system comprises means for recovery of the function blocked by expression of the blocking genes of the BCs. The invention is also related to a kit comprising a cloning vector having the BCs and TGI or alternatively a place for TGI to be inserted in. Segregating (two-insert) and inducible (one-insert) types of double RBF are disclosed. The two BCs are either similar or different in their DNA and RNA structures. The two BCs may encode same protein but promoters driving the blocking genes are different and/or the coding sequence of the blocking genes is modified by codon preference change. A system comprising two BCs having different blocking genes is also disclosed.

Abstract: The present invention is related to a recoverable block of function (RBF) system for controlling transgene escape and transgene containment in plants. The RBF-system comprises at least one blocking construct (BC), wherein the BC comprises as the blocking gene a nucleotide sequence or crtB gene encoding phytoene synthase, which is used alone or in combination with other BCs. The crtB gene expresses under the control of a germination specific promoter or a promoter constitutively repressed by a Recovering construct (RC) and blocks the germination of a transgenic seed. The blocked germination is recoverable by a user controlled intervention including addition of an effective amount of gibberellin and sugar and may involve so called recovering constructs (RCs), wherein the recovering gene may encode ent-copalyl diphosphate synthase or ent-kaurene synthase.

Abstract: The present invention provides a molecular mechanism for gene containment in sexually reproducing transgenic plants. The mechanism is achieved with a molecular construct comprising a blocking construct (BC) that is inserted fully or partially into an intron of a transgene of interest (TGI). The TGI encodes desired gene products, such as heterologous or homologous proteins, peptides or other useful products. The expression of the BC leads to block of at least one molecular or physiological function that is essential for development or reproduction of the transgenic plant. Thereby the BC expression leads to death or incapacity of sexual reproduction of the plant. Moreover, the mechanism comprises an externally applicable recovering tool to recover the functions blocked by the BC. The recovering tool may be a recovering construct (RC).

Abstract: The present invention is related to a family of novel spatiotemporally active Rubisco promoters (SEQ ID NO: 1, 2, 3) obtainable from light grown Brassica seedlings. Furthermore the invention is related to transgene expression in specific plant organs or at specific stages of plant development. DNA constructs and expression cassettes comprising at least one of the promoter sequences functionally fused in frame with genes encoding desired gene products are disclosed. Seeds from transformed homologous and heterologous plants and from subsequent generation of the transformed plants are collected and used for efficient production of desired gene products, especially in contained conditions.

Abstract: The present invention is related to a family of novel spatiotemporally active Rubisco promoters (SEQ ID NO: 1, 2, 3) obtainable from light grown Brassica seedlings. Furthermore the invention is related to transgene expression in specific plant organs or at specific stages of plant development. DNA constructs and expression cassettes comprising at least one of the promoter sequences functionally fused in frame with genes encoding desired gene products are disclosed. Seeds from transformed homologous and herterologous plants and from subsequent generation of the transformed plants are collected and used for efficient production of desired gene products, especially in contained conditions.

Abstract: The present invention is related to a family of novel spatiotemporally active Rubisco promoters (SEQ ID NO: 1, 2, 3) obtainable from light grown Brassica seedlings. Furthermore the invention is related to transgene expression in specific plant organs or at specific stages of plant development. DNA constructs and expression cassettes comprising at least one of the promoter sequences functionally fused in frame with genes encoding desired gene products are disclosed. Seeds from transformed homologous and herterologous plants and from subsequent generation of the transformed plants are collected and used for efficient production of desired gene products, especially in contained conditions.

Abstract: The invention is related to a method and a DNA construct for controlling transgene segregation in a sexually reproducing multicellular organism (SRMO), especially in such SRMOs which are susceptible of interbreeding with their cultured or wild-type relatives. The method and construct allows the farmer to reuse the transgenic crop without risk for escape of the transgene into the environment. The DNA construct is a recoverable block of function (RBF) system comprising at least one blocking construct (BC), and at least one means for recovering the blocked functions. The BC has a capacity of blocking at least one molecular or physiological function essential for the development and/or reproductive cycle of the SRMO. The BC is closely linked to the transgene of intereset (TGI). The blocked function is recoverable by an external treatment or intervention, which is optionally combined with one or more recovering constructs (RC).