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Rice serine hydroxymethyl transferase protein and function of coding gene thereof

A serine and gene encoding technology, applied in the field of plant genetic engineering, can solve problems such as difficult gene function research, and achieve the effect of broad application prospects

Inactive Publication Date: 2013-11-06
ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In rice, there are no related reports on serine hydroxymethyltransferase and its gene. Due to the lack of mutants of rice serine hydroxymethyltransferase gene, it is difficult to conduct in-depth functional research on the gene

Method used

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  • Rice serine hydroxymethyl transferase protein and function of coding gene thereof
  • Rice serine hydroxymethyl transferase protein and function of coding gene thereof
  • Rice serine hydroxymethyl transferase protein and function of coding gene thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Map-position cloning of embodiment 1, LZS gene

[0029] 1. Isolation and genetic analysis of the rice lzs mutant

[0030] The mutant was discovered from the tissue-cultured progeny of Zhonghua 11. From the second-leaf stage, the new leaves turned white, and the leaves gradually withered at the three-leaf stage (attached figure 1 ). In the T1 generation, the segregation ratio of wild-type phenotype plants and mutant plants was 35:13, in line with 3:1 (χ 2 c =0.032 0.05 =3.84) segregation relationship, the T1 generation seeds were harvested from a single plant, and 10 individual plants were randomly selected to plant the T2 generation, and 3 strains of the T2 generation strains showed wild-type phenotypes, no longer segregated, and 7 strains of plants The phenotypes were segregated, and the segregation ratio between wild-type phenotype plants and mutant plants was 3:1, suggesting that the mutant was controlled by a pair of recessive nuclear genes.

[0031] 2. Cloning...

Embodiment 2

[0040] Example 2: OsSHM1 functional complementation experiment

[0041] 1. Complementary carrier construction

[0042] Functional Complementary Fragment Select a DNA fragment of about 6.8 kb in total, including the entire ORF of the candidate gene LZS, the 5'UTR of about 2.1kb before the gene translation initiation codon ATG and the 3'UTR of about 660bp after the termination codon TAA, for the construction of a complementary vector . The amplification primers are:

[0043] 5'-AGCAC GTC GAC CAAAAGCAGGCTTACTGCCACAAAG-3' (SEQ ID NO: 19)

[0044] 3'-GTCCAC AGATCT CATATGCCTCTTACTCTCACAGTTG-3' (SEQ ID NO: 20)

[0045] The amplified product contains Sal I and Bgl II restriction sites for ligation to the pCAMBIA1301 vector. PCR amplification amplification system (50 μL): 25 μL of 2×Primer STAR GC buffer, 8 μL of dNTPs (2.5 mmol / L), 1 μL of each primer (10 mmol / L), 1 μL of TaKaRa Primer STAR Taq (5U / μL), DNA ( 50ng / μL) 1.5μL, sterile water 12.5μL. PCR amplification was perfor...

example 3

[0056] Example 3: Determination of photosynthetic pigment content

[0057] The fresh leaves of mutant lzs and wild type Zhonghua 11 with the same growth at the second leaf stage were taken respectively, and their chlorophyll content was measured. The results showed that due to the mutation of leaf color, the content of chlorophyll a, chlorophyll b and carotenoids of the mutant lzs at the second-leaf stage were significantly lower than those of the wild type, but the ratio of chlorophyll Chla / Chlb had no significant difference (attached Figure 8 )

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Abstract

The invention relates to a rice serine hydroxymethyl transferase protein and a function of a coding gene thereof, wherein the protein has an amino acid sequence shown in SEQ ID NO:2, and the gene for coding the protein has a nucleotide sequence shown in SEQ ID NO:1. The invention also discloses a plasmid containing the gene, a plant expression vector and a host cell. The invention also discloses a lethal method for the seedling stage of cultivars. The plant expression vector is used for converting plant cells, and the converted plant cells are cultured into plants. In the invention, the site of a new gene cloned in a mottle leaf lethal mutant in the seedling stage can be mutated to obtain a seedling-stage lethal phenotype. The invention has broad application prospects in hybrid seed production and purification as well as seed intellectual property protection.

Description

technical field [0001] The invention belongs to the field of plant genetic engineering. In particular, it relates to the function of rice serine hydroxymethyltransferase protein and its coding gene. Specifically, the present invention relates to a method of cloning a rice gene encoding serine hydroxymethyltransferase 1 (OsSHM1) gene by map-based cloning technology, and identifying the function of the gene by using a transgenic experiment. Background technique [0002] Serine Hydroxymethyltransferase (SHMT) is a pyridoxalase with pyridoxal 5'phosphate (PLP) as a coenzyme, which plays a very important role in the one-carbon metabolism and photorespiration of higher plants. There are two types of SHMT in eukaryotes, namely cytoplasmic serine hydroxymethyltransferase (cSHMT) and mitochondrial serine hydroxymethyltransferase (mSHMT). In prokaryotes, only cSHMT exists. The main functions of SHMT in plants are to provide Gly in the biosynthesis of proteins and purines, and to su...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/54C12N9/10C12N15/63C12N5/10A01H5/00
Inventor 汪得凯陶跃之严松刘合芹李素娟
Owner ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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