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Rice OsCS coded sequence and application thereof

A rice, coding technology, applied in the field of molecular biology, physiology and genetic engineering

Inactive Publication Date: 2007-10-24
FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Before the present invention was published, there was no disclosure or report of the rice OsCS sequence and its nucleic acid sequence mentioned in this patent application

Method used

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  • Rice OsCS coded sequence and application thereof
  • Rice OsCS coded sequence and application thereof
  • Rice OsCS coded sequence and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Cloning of Rice OsCS Gene

[0038] 1. Take the rice leaves grown in the greenhouse (25°C) (two leaves and one heart stage) and immediately put them in liquid nitrogen for cryopreservation. It should be noted that the materials selected in this experiment are not necessarily related to the selected rice varieties.

[0039] 2. Extraction of DNA

[0040]Take part of the tissue, grind it with a mortar, add it to a 50ml centrifuge tube filled with preheated lysate, keep it warm at 65°C for 40min, and centrifuge. RNaseA (0.5 μg / ml) was added to the supernatant, and the RNA was digested at 65°C for 1 h; the protein was pumped once with an equal volume of phenol / chloroform / isoamyl alcohol (25 / 24 / 1), the DNA was precipitated with ethanol, and washed twice with 70% ethanol. Repeat, and finally dissolve the DNA with TE solution. 0.8% agarose gel was used to check the quality of DNA.

[0041] 3. Cloning of the full-length gene

[0042] The degenerate primers were designed accor...

Embodiment 2

[0044] Sequence information and homology analysis of rice OsCS gene

[0045] The length of the novel rice OsCS gene of the present invention is 1425bp, and the detailed sequence is shown in SEQ ID NO.1. The polypeptide encoded by the gene consists of 475 amino acid residues, with a molecular weight of 52443.55 Daltons and an isoelectric point of 8.268. See SEQ ID NO.2 for the detailed sequence.

[0046] The coding region sequence of the full-length rice OsCS gene and its encoded protein sequence were analyzed by the BLAST program in the Non-redundant GeneBank+EMBL+DDBJ+PDB and Non-redundant GeneBank CDS translations+PDB+SwissPort+Superdate+PIR databases The homology of acid and protein was detected, and it was found that it has certain homology with the CS of carrot, tobacco, sugar beet, Arabidopsis and citrus. At the amino acid level, there is greater than 70% similarity (see Table 1). It can be seen from the above that the OsCS gene has high homology with the CS genes of c...

Embodiment 3

[0049] Copy number analysis of rice OsCS gene

[0050] A large amount of rice genomic DNA was extracted by CTAB method, 10 μg of DNA was digested with DraI, EcoRI and HindIII respectively, fragments were separated by 0.8% agarose gel, and the DNA was transferred to Hybond-N + fixed on a nylon membrane; using the rice OsCS gene as a probe to carry out Southern hybridization to identify its copy number in rice, the results showed that the OsCS gene was multi-copy.

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Abstract

The invention relates to a code sequence expressing citrate synthase in the rice and an application improving plant citric acid secretory and plant aluminium toxin immunity in the gene engineering technique field, which also relates to a recombinant expression vector of the gene, a conversion plant cell of expression vector, the transgenosis plant of the gene developed by the conversion cell. The gene expression in the plant improves the citric acid secretory quantity of the transgenosis plant, which enhances the aluminium toxin immunity.

Description

technical field [0001] The invention belongs to the technical fields of molecular biology, physiology and genetic engineering, and in particular relates to the cloning, copy number analysis and gene expression pattern analysis of a citrate synthase (rice citrate synthase, OsCS) expressed in rice , the construction of transgenic vectors, plant transformation, the acquisition of transgenic plants and the identification of corresponding physiological and biochemical characteristics. Background technique [0002] Nearly 40% of the world's cultivated soil is eroded by acid rain. Among them, aluminum (Al) toxicity is a key factor limiting crop yield in acidic soils. The use of genetic engineering to breed good varieties adapted to poor soil has a certain application potential. The organic acids secreted by roots are of certain importance to the absorption of mineral nutrients by plants. Most importantly, the secretion of organic acids improves the resistance of plants to alumin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/52C12N9/00C12N15/82C12Q1/68
Inventor 明凤张珊珊沈大棱
Owner FUDAN UNIV
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