Transcription in plants and bacteria

Claim 2

In a method of genetically modifying a cell comprising the steps of: (a) transforming a prokaryotic cell with a DNA molecule comprising a promoter region 5' from a foreign structural gene, wherein said promoter region causes transcription of said structural gene in prokaryotes, and (b) detecting expression of said gene in a resultant prokaryotic strain, the improvement comprising transforming said prokaryotic cell with a DNA molecule comprising a T-DNA gene promoter region 5' from a foreign structural gene, said foreign structural gene not being under control of said promoter in nature, and said T-DNA region being capable of causing transcription of said gene in plant cells as well as prokaryotes.

Claim 3

In a method for transforming transformable plant cell with a DNA molecule comprising a promoter region 5' from a foreign structural gene, wherein said promoter region causes transcription of said structural gene in plant cells by: (a) transforming a transformable plant cell with said DNA molecule; and (b) detecting expression of said gene in a resultant plant tissue, the improvement comprising utilizing the method of claim 1 to first transform a prokaryotic cell with said DNA molecule and detect expression of said gene therein.

Claim 4

A method according to claim 2 wherein said promoter region is hybridizable under conditions of appropriate stringency to the 1450Tx gene promoter.

Claim 5

A method according to claim 2 wherein said promoter region is the T-DNA 1450bTx gene promoter.

Claim 6

A method according to claim 2 wherein said structural gene encodes a resistance to an antibiotic selected from the group consisting of kanamycin, neomycin, G418 and an analog thereof.

Claim 7

A method according to claim 5 wherein said structural gene encodes neomycin phosphotransferase II from Tn5.

Claim 8

A method according to claim 2 wherein said DNA molecule is linked to a replicon that cannot be maintained independently in a bacterium of the family Rhizobiaceae.

Claim 9

A method according to claim 2 wherein said DNA molecule is linked to a replicon that can be maintained independently in a bacterium of the family Rhizobaeceae.

Claim 10

A method according to claim 2 wherein said DNA molecule is linked to one or more of the repetitive sequences selected from the group consisting of T L LB(A), T L RB(B), T R LB(C) and T R RB(D).

Claim 11

A method according to claim 2 wherein said structural gene is ligated to the left end of the DNA fragment d 2 of the TR section of T-DNA.

Claim 12

A method according to claim 10 wherein said structural gene is ligated to the right end of the T-DNA fragment b 4 of the TR section of T-DNA.

Claim 13

A DNA vector comprising (a) a T-DNA region containing structural elements essential for transfer of said T-DNA region into a dicot plant genome; and (b) one or more T-DNA promoter regions 5' from a foreign structural gene inserted in said T-DNA region and capable of conferring an identifiable phenotype, said promoter region being a T-DNA gene promoter region that causes transciption both in prokaryotes and in dicot plant cells, said foreign structural gene being a gene which is not under the control of said promoter in nature, said promoter region and foreign strucutral gene being inserted in said T-DNA region so that the transfer activity of the T-DNA is not disrupted, and said foreign structural gene being the sole means for said vector to confer said identifiable phenotype to a bacterial strain transformed by said vector.

Claim 14

A DNA vector comprising (a) a T-DNA region containing structural elements essential for transfer of said T-DNA region into a dicot plant genome; and (b) one or more T-DNA promoter regions 5' from a foreign structural gene inserted in said T-DNA region and capable of conferring an identifiable phenotype, said promoter region being a T-DNA gene promoter region that causes transciption both in prokaryotes and in dicot plant cells, said foreign structural gene being a gene which is not under the control of said promoter in nature, said promoter region and foreign strucutral gene being inserted in said T-DNA region so that the transfer activity of the T-DNA is not disrupted, and said foreign structural gene being the sole means for said vector to confer said identifiable phenotype to a bacterial strain transformed by said vector.

Claim 15

A vector according to claim 12 wherein said promoter region is hybridizable under conditions of appropriate stringency to the 1450bTx gene promoter.

Claim 16

A vector according to claim 12 wherein said promoter region is the T-DNA 1450bTx gene promoter.

Claim 17

A vector according to claim 12 wherein said structural gene confers an identifiable phenotype in a plant cell transformed to contain said structural gene.

Claim 18

A vector according to claim 15 wherein said identifiable phenotype is a resistance to an antibiotic selected from the group consisting of kanamycin, neomycin, G418 and an analog thereof.

Claim 19

A vector according to claim 16 wherein said structural gene encodes neomycin phosphotransferase II from Tn5.

Claim 20

A vector according to claim 17 wherein said vector is selected from the group consisting of pRK290K and-1 and functionally equivalent pRK290 based shuttle vectors.

Claim 21

A bacterial strain comprising the vector of claim 18.

Claim 22

A vector according to claim 12, further comprising a replicon that cannot be maintained independently in a bacterium of the family Rhizobiaceae.

Claim 23

A vector according to claim 12, further comprising a replicon that can be maintained independently in a bacterium of the family Rhizobiaceae.

Claim 24

A vector according to claim 12, further comprising one or more of the repetitive sequences selected from the group consisting of T L LB(A), T L RB(B). T R LB(C) and T R RB(D).

Claim 25

A vector according to claim 12 wherein said structural gene is ligated to the left end of the DNA fragment d 2 of the TR section of T-DNA.

Claim 26

A vector according to claim 23 wherein said structural gene is ligated to the right end of the DNA fragment b 4 of the TR section of T-DNA.