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Jatropha curcas phytoene synthetase gene sequence and application thereof in plants

A technology of phytoene and synthetase, applied in the fields of molecular biology and genetic engineering

Inactive Publication Date: 2012-05-23
FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Before the present invention was published, there was no publication or report of the cDNA sequence encoding Jatropha curcas phytoene synthase mentioned in this patent application

Method used

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  • Jatropha curcas phytoene synthetase gene sequence and application thereof in plants
  • Jatropha curcas phytoene synthetase gene sequence and application thereof in plants
  • Jatropha curcas phytoene synthetase gene sequence and application thereof in plants

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] Cloning of PSY Protein Gene of Jatropha curcas

[0063] 1. Tissue separation (isolation)

[0064] The seeds of Jatropha curcas come from Panzhihua, Sichuan Province. After the seeds of Jatropha curcas are collected, they are stored in the laboratory.

[0065] 2. RNA isolation (RNA isolation)

[0066] Peel off the seed coat of Jatropha curcas seeds, take out the seed kernels, grind them with a mortar, add liquid nitrogen, grind them into powder, take 100mg and transfer them to 1.5mL EP tubes, and extract total RNA (two-step lysis method) . The quality of total RNA was identified by formaldehyde denaturing gel electrophoresis, and then the RNA content was determined on a spectrophotometer.

[0067]3. Cloning of Full-length cDNA

[0068] According to the conserved amino acid sequence of PSY in some plants, design degenerate primers, using the principle of homologous gene cloning, SMART TM RACE cDNA amplification method (Clonetech kit) for full-length cDNA cloning is ...

Embodiment 2

[0079] Sequence information and homology analysis of PSY protein gene of Jatropha curcas

[0080] The full-length cDNA of the new Jatropha curcas PSY protein of the present invention is 1755bp in length, and the detailed sequence is shown in SEQ ID NO.1, wherein the open reading frame is located at nucleotides 78-1364. The amino acid sequence of Jatropha curcas PSY protein was deduced according to the full-length cDNA, with a total of 429 amino acid residues, a molecular weight of 48068.52, and a pI of 8.96. See SEQ ID NO.2 for the detailed sequence.

[0081] The full-length cDNA sequence of the Jatropha curcas PSY protein and its encoded protein were nucleotide aligned in the Non-redundant GenBank+EMBL+DDBJ+PDB and Non-redundant GenBank CDStranslations+PDB+SwissProt+Superdate+PIR databases using the BLAST program. Protein homology search, the results found that the nucleotide sequence of Jatropha curcas PSY gene and other plant nucleotide sequences such as sweet orange (C.si...

Embodiment 3

[0082] The structural characteristic analysis of embodiment 3 Jatropha curcas PSY protein

[0083] 1. Domain analysis of Jatropha curcas PSY protein

[0084] The amino acid sequence of the Jatropha curcas PSY protein is retrieved in the NCBI database (the website is: http: / / www.ncbi.nlm.nih.gov / Blast.cgi), and the results are shown in image 3 .

[0085] In the amino acid sequence, there is a catalytic site of isopentenyl pyrophosphate synthase (Trans-Isoprenyl Diphosphate Synthases, Trans_IPPS) (i.e. amino acid residues 137-404 from the amino terminal (N terminal) of SEQ ID.1 in the sequence listing .

[0086] 2. Analysis of signal peptide of Jatropha curcas PSY protein

[0087] Plant carotenoid synthases are encoded by nuclear genes, and the codes are translated to form precursors (precursors) with transit peptide sequences at the N-terminal. Transit peptides have the function of transporting plant carotenoid synthases to plastids. After processing or translocation, they...

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Abstract

The invention relates to the technical fields of molecular biology and genetic engineering and relates to a JcPSY protein for expressing the Jatropha curcas phytoene synthetase and a nucleotide sequence thereof. Due to the use of the Jatropha curcas PSY protein, substances interacting with the Jatropha curcas PSY protein, or receptors, inhibitors or antagonists, and the like can be screened by various conventional screening methods. The Jatropha curcas PSY protein genes can be used for improving the beta-carotene content in the plants and improving the economic values of the plants through the genetic engineering technology, and have obvious market application prospects.

Description

technical field [0001] The invention relates to the technical fields of molecular biology and genetic engineering. Specifically, the present invention relates to a method for expressing JcPSY protein (Jatropha curcas phytoenesynthase, JcPSY) and its nucleic acid sequence in Jatropha curcas. Background technique [0002] It is known that the biosynthesis of β-carotene mainly occurs in higher plants, algae, fungi and bacteria, and β-carotene cannot be synthesized de novo in animals. β-carotene is the precursor of vitamin A (Vitamin A), which can be converted into vitamin A in the human body as needed. It has dual functions of nutrition and coloring, and it is an excellent and nutritious food additive (food additives) of Class A identified by the Joint Expert Committee on Food Additives of the Food and Agriculture Organization of the United Nations (FAO) and the World Health Organization (WHO). In recent years, more and more medical studies have shown that β-carotene can resi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/52C12N9/00C12N15/63C12N1/19C12N5/10C12N15/82C07K16/40C12Q1/68C12N15/11
Inventor 林娟侯嵘周选围
Owner FUDAN UNIV
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