# US 2023/0407318 A1 · TEA PLANT CsVAAT3 GENE AND USE THEREOF

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- Number: US 2023/0407318 A1
- Document ID: US20230407318A1
- Status: Published application
- Kind: A1
- Application number: 18/305,944
- Filed: 2023-04-24
- Published: 2023-12-21
- Earliest claimed: 2022-04-24
- Assignees: Anhui Agricultural University
- Applicants: Anhui Agricultural University
- Inventors: Tianyuan YANG; Junjie WANG; Xiaojuan FAN; Zhaoliang ZHANG; Xiaohong LIAN; Xinpeng ZHAO; Xiaocao LUO
- Web page: https://patentgrep.com/patent/US20230407318A1

## CPC

- C12N 15/81: for yeasts
- C12N 1/16: Yeasts; Culture media therefor
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

The present disclosure provides a tea plant CsVAAT3 gene and use thereof. The tea plant CsVAAT3 gene has a nucleotide sequence shown in SEQ ID NO: 1 in sequence listing. A protein coded by the tea plant CsVAAT3 gene has an amino acid sequence shown in SEQ ID NO: 2 in the sequence listing. An expression pattern of CsVAAT3 in tea plant is significantly positively correlated with the nitrogen level in tea roots. The CsVAAT3 is highly expressed in tea roots. A pDR196-CsVAAT3 plasmid constructed from this gene is transformed into a vacuolar amino acid uptake-deficient yeast strain ypq2, which can restore the growth ability of the yeast mutant on a high-concentration theanine medium. Cloning of the gene is beneficial to analysis of a molecular mechanism of theanine storage in tea roots, providing an important target gene resource and a theoretical basis for cultivation of new tea cultivars with high theanine content.

## Claims

### Claim 1

A tea plant CsVAAT3 gene, wherein the tea plant CsVAAT3 gene is a tonoplast-localized transporter gene, and the tea plant CsVAAT3 gene has a nucleotide sequence shown in SEQ ID NO: 1 in sequence listing.

### Claim 2

(canceled)

### Claim 3

A tea plant expression vector pDR196-EGFP-CsVAAT3, wherein the expression vector is obtained by digesting a fragment shown in SEQ ID NO: 1 into a pDR196-EGFP vector.

### Claim 4

(canceled)

### Claim 5 (depends on 1)

A method for transporting theanine into a yeast vacuole in yeast, comprising the following steps:
cloning the tea plant CsVAAT3 gene according to claim 1;
constructing a tea plant expression vector; and
transforming the tea plant CsVAAT3 into a yeast mutant, wherein the yeast mutant is a high-concentration theanine-sensitive strain, and the yeast mutant suffers from inhibited growth under a high-concentration theanine environment.

### Claim 6 (depends on 5)

The method for transporting theanine into a yeast vacuole in yeast according to claim 5, wherein the tea plant expression vector is pDR196-EGFP-CsVAAT3.

### Claim 7

(canceled)