Jatropha curcas farnesyl pyrophosphate synthase protein encoding sequence and application thereof in plants

A Jatropha curcas, nucleotide sequence technology, applied in the fields of genetic engineering and molecular biology

Inactive Publication Date: 2010-08-11
FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] Before the present invention was published, there was no disclosure or report of the cDNA sequence encoding Jatropha curcas farnesyl pyrophosphate synthase mentioned in this patent application

Method used

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  • Jatropha curcas farnesyl pyrophosphate synthase protein encoding sequence and application thereof in plants
  • Jatropha curcas farnesyl pyrophosphate synthase protein encoding sequence and application thereof in plants
  • Jatropha curcas farnesyl pyrophosphate synthase protein encoding sequence and application thereof in plants

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0137] Cloning of JcFPS Protein Gene of Jatropha curcas

[0138] 1. Tissue separation (isolation)

[0139] The seeds of Jatropha curcas come from Panzhihua, Sichuan Province. After the seeds of Jatropha curcas are collected, they are stored in the laboratory.

[0140] 2. RNA isolation (RNA isolation)

[0141] 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.

[0142] 3. Cloning of Full-length cDNA

[0143] According to the amino acid conserved sequence of some plant FPS, design annexation primers, use the principle of homologous gene cloning, and use the SMARTTMRACE cDNA amplification method (Clonetech kit) to carry out full-length cD...

Embodiment 2

[0153] Sequence Information and Homology Analysis of JcFPS Protein Gene of Jatropha curcas

[0154] The length of the full-length cDNA of the new Jatropha curcas JcFPS protein of the present invention is 1029bp, and the detailed sequence is shown in SEQ ID NO.1, wherein the open reading frame is located at nucleotides 221-1246. The amino acid sequence of Jatropha curcas JcFPS protein was deduced according to the full-length cDNA, with a total of 381 amino acid residues, a molecular weight of 43784.45, and a pI of 5.54. See SEQ ID NO.2 for the detailed sequence.

[0155] The full-length cDNA sequence of the Jatropha curcas JcFPS 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, it was found that it has 87% homology (attached table 2) with Brasiliensis HbFPS gene (AB294712) at the nucleotide level; AA...

Embodiment 3

[0156] The structural characteristic analysis of embodiment 3 Jatropha curcas FPS protein

[0157] 1. Domain analysis of Jatropha curcas FPS protein

[0158] The amino acid sequence of the Jatropha curcas FPS protein is retrieved in the NCBI database (URL: http: / / www.ncbi.nlm.nih.gov / Blast.cgi), and the results show that:

[0159]In the amino acid sequence, there is the catalytic site of Trans-Isoprenyl Diphosphate Synthases (Trans-Isoprenyl Diphosphate Synthases, Trans IPPS) (ie, amino acid 69-303 from the amino terminal (N terminal) of SEQ ID.1 in the sequence listing Residues.

[0160] 2. Hydrophobicity analysis of Jatropha curcas FPS protein

[0161] Analysis of the mature FPS after shearing of the transit peptide shows that, on the whole, the hydrophilic amino acids are evenly distributed in the entire peptide chain, and the entire polypeptide chain is hydrophilic without obvious hydrophobic regions. The hydrophobicity analysis chart of the present invention is establi...

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Abstract

The invention belongs to the technical fields of molecular biology and genetic engineering, and provides an encoding sequence for encoding cDNA of Jatropha curcas farnesyl pyrophosphate synthase. The farnesyl pyrophosphate synthase catalyzes isopentenyl pyrophosphate and dimethylallyl pyrophosphate to carry out continuous condensation reaction to form farnesyl pyrophosphate, is key enzyme for metabolic pathway of Jatropha curcas isoprene, and contributes to reducing the content of phorbol esters and precursor thereof in the Jatropha curcas. Due to the use of the Jatropha curcas protein, substances interacting with the Jatropha curcas protein, or receptors, inhibitors or antagonists, and the like can be screened by various conventional screening methods. The encoding sequence for encoding the cDNA of the Jatropha curcas farnesyl pyrophosphate synthase has great application prospects.

Description

technical field [0001] The invention belongs to the technical fields of molecular biology and genetic engineering. Specifically, the present invention relates to a jc-fps protein (Jatropha curcas farnesyl pyrophosphate synthase protein, Jatropha curcas farnesyl pyrophosphate synthase, JCFPS) expressed in Jatropha curcas and its nucleic acid sequence. Background technique [0002] Jatropha curcas, also known as Jatropha, is a Euphorbiaceae plant distributed in Sichuan, Yunnan, Guangxi and other places in my country. In recent years, Jatropha curcas has been widely developed as a new type of bioenergy. In addition, Jatropha curcas is also used in many fields such as pharmaceuticals, biological pesticides, wasteland management, pharmaceuticals, feed, and fertilizers. Jatropha curcas contains a variety of toxin components, and the more toxic components are jatropha curcas toxin and phorbol ester. Phorbol ester (12-deoxy-16-hydroxyphorbol, 12-deoxy-16-hydroxyphorbol) is a diter...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/54C12N9/10C12N15/63C12N1/19C12N5/10C12N15/82C07K16/40C12Q1/68
Inventor 林娟侯嵘周选围
Owner FUDAN UNIV
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