# US 7,608,758 B2 · Plant system for comprehensive gene function analysis with the use of full-length cDNA

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- Number: US 7,608,758 B2
- Document ID: US7608758B2
- Status: Granted
- Kind: B2
- Application number: 10/488,087
- Filed: 2002-08-29
- Published: 2004-10-14
- Granted: 2009-10-27
- Earliest claimed: 2001-08-31
- Assignees: Riken
- Inventors: Minami Matsui; Takanari Ichikawa; Miki Nakazawa; Motoaki Seki; Miki Fujita; Kazuo Shinozaki
- Web page: https://patentgrep.com/patent/US7608758B2

## CPC

- C12N 15/8216: {Methods for controlling, regulating or enhancing expression of transgenes in plant cells}
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

A gene causative of a phenotype is easily specified and thus gene functions are comprehensively analyzed. A method for analyzing a gene function includes (a) infecting a plant group with a full-length cDNA library containing an expression regulatory sequence to introduce the cDNA into the plants; (b) selecting a plant complying with desired selection requirements from the plants into which the cDNA has been introduced; (c) isolating the cDNA from the selected plant; and (d) re-introducing the thus isolated cDNA into a plant of the same species as the plant into which the cDNA has been introduced, to reconfirm a phenotype according to the selection requirements.

## Claims

### Claim 1

A method for analyzing a gene function comprising the steps of:
(a) preparing a normalized full-length cDNA library containing an expression regulatory sequence by cloning full-length cDNAs from an organism into vectors, sequencing the 5′ and 3′ terminal sequences of the full-length cDNA clones, selecting all of the full-length cDNA clones that are different from each such that there is no overlapping, and mixing the selected full-length cDNA clones in equivalent amounts;
(b) infecting a plant group with the normalized full-length cDNA library to introduce the cDNA into the plants;
(c) selecting a plant complying with desired selection requirements from the plant group having the cDNA introduced thereinto;
(d) isolating the cDNA from the selected plant; and
(e) re-introducing the isolated cDNA into a plant of the same species as the plant into which the cDNA has been introduced to reconfirm a phenotype according to the selection requirements.

### Claim 2 (depends on 1)

The method according to claim 1, wherein the expression regulatory sequence is a constitutive expression promoter, an inducible promoter, or a combination thereof.

### Claim 3 (depends on 1)

The method according to claim 1, wherein the full-length cDNA library is introduced into Agrobacterium.

### Claim 4 (depends on 1)

The method according to claim 1, wherein the cDNA is introduced into the plant by infection with Agrobacterium.

### Claim 5 (depends on 1)

The method according to claim 1, wherein the selection requirements are based on change of phenotype for stress resistance and/or morphogenesis.

### Claim 6 (depends on 5)

The method according to claim 5, wherein the stress is at least one form of stress selected from the group consisting of oligotrophic stress, drought stress, temperature stress, strong light stress, ultraviolet stress, salt stress, air pollution stress, agricultural chemical stress, oxidization stress, heavy metal stress, disease damage stress, and hormone stress.