Separated eggplant Cu/ZnSOD protein as well as preparation and applications thereof

A protein and eggplant technology, which is applied to the separated eggplant Cu/ZnSOD protein and its preparation and application fields, can solve the problem of no encoded protein, etc., and achieve the effect of improving salt tolerance and great application value.

Inactive Publication Date: 2017-10-10
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no relevant literature report on eggplant Cu / ZnSOD gene and its encoded protein

Method used

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  • Separated eggplant Cu/ZnSOD protein as well as preparation and applications thereof
  • Separated eggplant Cu/ZnSOD protein as well as preparation and applications thereof
  • Separated eggplant Cu/ZnSOD protein as well as preparation and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] The cloning of embodiment 1, eggplant Cu / ZnSOD gene

[0040] 1. Acquisition of plant material

[0041] The plant material used in this experiment is the excellent eggplant germplasm resource A. spp. The experimental materials were cultivated in artificial plastic greenhouses in the Aerospace Breeding Base of Pujiang Town, Minhang District, Shanghai. Seedlings are raised, grown and fruited under natural conditions. The leaves of eggplant were collected for RNA extraction.

[0042] 2. Extraction of RNA

[0043] Total RNA was extracted by the TRIzol method (TRIzol was purchased from Sangon Bioengineering (Shanghai) Co., Ltd.). The integrity of RNA was identified by formaldehyde denaturing gel electrophoresis, and then the purity and concentration of RNA were determined on a spectrophotometer (Thermo Scientific NANODROP 1000 Spectrophotometer).

[0044] 3. Full-length cloning of genes

[0045] A pair of degenerate primers was designed based on the conserved sequence o...

Embodiment 2

[0050] Example 2, Sequence Information and Homology Analysis of Eggplant Cu / ZnSOD Gene

[0051] The full-length CDS open reading frame sequence of the new eggplant Cu / ZnSOD gene obtained in the present invention is 459 bp, and the detailed sequence is shown in SEQ ID NO.1. According to the CDS open reading frame sequence, the amino acid sequence of eggplant Cu / ZnSOD is known, with a total of 152 amino acid residues, a molecular weight of 15.25Kd (kilodaltons), and a theoretical isoelectric point (pI) of 5.47. The sequence is as shown in SEQ ID NO. 2, specifically:

[0052] MVKAVAVLNSSEGVSGTIFFTQEADVPTTVSGNISGLKPGLHGFHVHALGDTTNGCMSTGPHFNPAGKDHGAPEDELRHAGDLGNITAGEDGTASFTITDKQIPLTGSQSIIGRAVVVHADPDDLGKGGHELSKSTGNAGGRIACGIIGLQG.

[0053] The CDS open reading frame sequence of eggplant Cu / ZnSOD and the amino acid sequence of its encoded protein were obtained by BLAST program in Non-redundant GenBank+EMBL+DDBJ+PDB and Non-redundant GenBank CDS translations+PDB+SwissProt+Superdate+PI...

Embodiment 3

[0055] Embodiment 3, expression of eggplant Cu / ZnSOD gene in different tissues

[0056] 1. Acquisition of plant material

[0057] The eggplant roots, stems, leaves, flowers, pericarp, and pulp were collected at the ripening stage of the fruit, and the samples were wrapped in aluminum platinum paper and immediately put into liquid nitrogen, and then transferred to a -80°C ultra-low temperature refrigerator for storage until use.

[0058] 2. Extraction of RNA

[0059] Total RNA was extracted by the TRIzol method (TRIzol was purchased from Sangon Bioengineering (Shanghai) Co., Ltd.). The integrity was detected by ordinary agarose gel electrophoresis (gel concentration 1.2%; 0.5×TBE electrophoresis buffer; 150v, 15min). The maximum rRNA brightness in the electrophoresis strip should be 1.5-2.0 times the brightness of the second rRNA, otherwise it indicates the degradation of the rRNA sample. RNA with good purity, A 260 / A 280 and A 260 / A 230 About 2.0 or so. The OD value ...

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Abstract

The invention belongs to the field of biotechnologies, and particularly relates to separated eggplant Cu / ZnSOD protein as well as preparation and application of the separated eggplant Cu / ZnSOD protein. The amino acid sequence of the separated eggplant Cu / ZnSOD protein is shown in SEQ ID NO.2. Aiming at the current situation that the existing research foundation for eggplant is weak, the key gene eggplant Cu / ZnSOD related to stress resistance is successfully separated and cloned for the first time, thus a theoretical basis is provided for improving the stress-resistance traits of plants by utilizing the genetic engineering technology and obtaining medicines or foods with high oxidation resistance in the future, and the application value is great.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to an isolated eggplant Cu / ZnSOD protein and its preparation and application. Background technique [0002] SOD is a metalloenzyme widely present in eukaryotes and aerobic prokaryotes, it is not only the first line of defense against reactive oxygen species, but also the first one involved in superoxide anion Cleared enzyme protein. superoxide anion are the main free radicals in living organisms; in many cases It is harmful to the body, and it is also one of the causes of aging. SOD is an important class of antioxidant enzymes that scavenge oxygen free radicals, it can catalyze disproportionation reaction to generate O 2 and H 2 o 2 ;H 2 o 2 Under the action of catalase (CAT), non-toxic H 2 O and O 2 , so as to play an anti-aging effect. [0003] Plant cells can produce reactive oxygen species under normal metabolic activities and stress conditions. In recen...

Claims

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Application Information

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Patent Type & AuthorityApplications(China)
IPC IPC(8): C12N9/02C12N15/53C12N15/70C12N1/21
CPCC12N9/0089C12Y115/01001
Inventor刘杨徐龙任丽蒋明敏刘卫
OwnerSHANGHAI JIAO TONG UNIV