Abstract: A method is disclosed for making more efficient the particle-mediated germ line genetic transformation of bean species such as soybean. After a particle-mediated transformation event, in the absence of a selectable marker gene, relatively large numbers of plants must be regenerated to find the relatively low likelihood germ line transformation events which have occurred. It has been discovered that using in the transformation process a marker gene linked to the gene of interest, and by excising a segment of the stem of the shoot during the regeneration process and assaying the segment for the marker gene, certain patterns or phenotypes can be identified in the stem segment which are associated with an increased frequency of germ line transformation events. As the plants are regenerated, other indices of gene expression, at the first trifoliate leaf stage and at the third or fourth trifoliate leaf stage, also serve as markers of the likelihood of germ line transformation.

Abstract: A method of genetically engineering a fiber-producing plant is disclosed. This method first comprises the step of constructing a plant expression vector that comprises a protein coding sequence and a DNA sequence capable of promoting gene expression in fiber cells, wherein the DNA sequence is homologous to a sequence selected from the group consisting of 12 different genomic sequences. The method next involves introducing the expression vector into a fiber-producing plant wherein the protein coding sequence is expressed in the fiber cells of the fiber producing plant. A method of obtaining a DNA sequence capable of promoting gene expression in fiber cells is also disclosed.

Abstract: A fiber-producing plant comprising in its genome a heterologous genetic construct is disclosed. This genetic construct comprises a fiber-specific promoter and a coding sequence encoding a plant peroxidase. Preferably, the coding sequence is for cotton peroxidase. Seeds of the plant containing this genetic construct and plant cells containing this construct are also disclosed.

Abstract: A plant expression vector is constructed to cause the expression of an amino-terminal portion of the Bacillus thuringiensis delta-endotoxin gene in plant cells and the vector is used to create transgenic plants expressing the toxin. A truncated form of the toxin is used, with carboxy-terminal prolines added for stability. A translational enhancer sequence derived from the untranslated leader sequence from the mRNA of the coat protein gene of alfalfa mosaic virus coat protein gene is placed between a promoter and the toxin gene to increase translational efficiency. The transgenic plants produced are toxic to Lepidopteran pests and can transmit that trait to their progeny by normal Mendelian inheritance.

Abstract: A gas driven apparatus for accelerating particles coated with a genetic material into a target comprises a reservoir for releasably retaining a gas at a sufficiently high pressure to detach the particles from the surface of a sample cartridge and to carry the particles through the apparatus toward the target. When leaving the apparatus, the particles entrained in the gas stream pass through a substantially conical exit nozzle which causes the pattern of distribution of the particles to greatly expand. Methods for using the apparatus and for preparing the sample cartridges are also described.

Abstract: An apparatus is disclosed for the genetic transformation of organisms by accelerated particle mediated transformation. Foreign genes are introduced into cells by coating on carrier particles which are physically accelerated into the cells by positioning the carrier particles on the external surface of a carrier ribbon which is wound on a cartridge, the carrier ribbon having an exposed portion. The carrier particles on the exposed portion of the ribbon are displaced and accelerated toward an exit port by a high pressure stream of helium gas. By rotating the ribbon, a continuous supply of carrier particles can be produced and hence large target areas can be transformed without the need to replace the particle carrier. Near the exit port, the gaseous stream is diverted through use of the Coanda effect to divert the gas stream away from a target area. The carrier particles, being much heavier than the gas, continue toward and into the target cells.

Abstract: A method of genetically engineering a fiber-producing plant is disclosed. This method first comprises the step of constructing a plant expression vector that comprises a protein coding sequence and a DNA sequence capable of promoting gene expression in fiber cells, wherein the DNA sequence is homologous to a sequence selected from the group consisting of 12 different genomic sequences. The method next involves introducing the expression vector into a fiber-producing plant wherein the protein coding sequence is expressed in the fiber cells of the fiber producing plant. A method of obtaining a DNA sequence capable of promoting gene expression in fiber cells is also disclosed.

Abstract: An apparatus for acceleration of carrier particles into a target organism which may be used for rapid, sequential particle acceleration procedures comprises a holder for removable mounting cartridges. A first removable mounting cartridge detachably holds a carrier sheet coated with carrier particles which have themselves been coated with biological material. A second removable mounting cartridge comprises a retaining screen. The mounting cartridges may be inserted into the holder before particle acceleration and replaced quickly afterwards, thereby readying the apparatus for another particle acceleration. Convenient replacement of these consumable items eliminates the need to disassemble the apparatus between each particle acceleration procedure.

Abstract: A method is disclosed for making more efficient the particle-mediated germ line genetic transformation of bean species such as soybean. After a particle-mediated transformation event, in the absence of a selectable marker gene, relatively large numbers of plants must be regenerated to find the relatively low likelihood germ line transformation events which have occurred. It has been discovered that using in the transformation process a marker gene linked to the gene of interest, and by excising a segment of the stem of the shoot during the regeneration process and assaying the segment for the marker gene, certain patterns or phenotypes can be identified in the stem segment which are associated with an increased frequency of germ line transformation events. As the plants are regenerated, other indices of gene expression, at the first trifoliate leaf stage and at the third or fourth trifoliate leaf stage, also serve as markers of the likelihood of germ line transformation.

Abstract: A method of genetically engineering a fiber-producing plant is disclosed. This method first comprises the step of constructing a plant expression vector that comprises a protein coding sequence and a DNA sequence capable of promoting gene expression in fiber cells, wherein the DNA sequence is homologous to a sequence selected from the group consisting of 12 different genomic sequences. The method next involves introducing the expression vector into a fiber-producing plant wherein the protein coding sequence is expressed in the fiber cells of the fiber producing plant. A method of obtaining a DNA sequence capable of promoting gene expression in fiber cells is also disclosed.

Abstract: Transgenic cotton plants have been created which expressed an immobilized protein in the cotton fiber cells. The cotton fiber can be recovered from such transgenic cotton plants and then used as a substrate for fixing immobilized protein for use in industrial or laboratory processes. Using an enzyme expressed inside cotton fiber, it is possible to fabricate a convenient reaction column by simply packing the cotton fiber carrying the enzyme into a column and passing substrate therethrough.

Abstract: An apparatus for the particle mediated genetic transformation of organisms in vivo has two parts, a support unit and a hand unit. The hand unit is of a convenient hand manipulable size so that it can be placed readily against the organism the cells of which are to be transformed. The hand unit is connected to the support unit by a flexible umbilical so that the hand unit can be placed where desired. The apparatus is particularly well adapted for the convenient transformation of somatic cells of whole animals or humans.

Abstract: A method for determining the developmental stage of a tumor is disclosed. The method comprises the steps of isolating tumor cells, dividing the cells into two samples, transducing one sample with a reporter gene, transducing the other sample with both a reporter gene and a staging gene (wherein the staging gene corresponds to a gene that is differentially expressed in neoplastic and normal cells and is chosen by its ability to be predictive of a certain stage of neoplastic development), analyzing both samples for the level of expression of the reporter gene and comparing the expression level in the samples. In a particularly advantageous embodiment of the invention, the staging gene is JR-ras and the reporter gene is lux.

Abstract: An apparatus for the particle mediated genetic transformation of organisms in vivo has two parts, a support unit and a hand unit. The hand unit is of a convenient hand manipulable size so that it can be placed readily against the organism the cells of which are to be transformed. The hand unit is connected to the support unit by a flexible umbilical so that the hand unit can be placed where desired. The apparatus is particularly well adapted for the convenient transformation of somatic cells of whole animals or humans.

Abstract: An apparatus is disclosed for the genetic transformation of organisms by accelerated particle mediated transformation. Foreign genes are introduced into cells by coating on carrier particles which are physically accelerated into the cells by positioning the carrier particles on a carrier sheet which is accelerated by a device which produces a shock wave. The treated cells are recovered, and a portion of them will contain in their genome the foreign gene. The procedure may be used to create genetically engineered organisms of many types.