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Application of PSA3 (photosystem I assembly 3) protein in assembly of PSI (photosystem I) and maintaining of PSI stability

A protein and stability technology, applied in the application field of PSA3 protein in participating in the assembly of PSI and maintaining the stability of PSI, can solve the problems of poorly understood biogenesis

Inactive Publication Date: 2017-09-22
INST OF BOTANY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In the prior art, little is known about the biogenesis of PSI

Method used

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  • Application of PSA3 (photosystem I assembly 3) protein in assembly of PSI (photosystem I) and maintaining of PSI stability
  • Application of PSA3 (photosystem I assembly 3) protein in assembly of PSI (photosystem I) and maintaining of PSI stability
  • Application of PSA3 (photosystem I assembly 3) protein in assembly of PSI (photosystem I) and maintaining of PSI stability

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0092] Example 1. Function discovery of PSA3 protein

[0093] The inventors of the present invention constructed a Mu-transposon-induced maize mutant library, from which the mutant strains (plants with yellowing phenotype) that could not undergo photosynthesis were screened, and the Mu-transposon insertion and phenotype were identified by a deep sequencing method Co-isolated, several insertion mutations were identified. A mutant strain has an insertion site near the start codon of a specific gene, and the mutant strain is named mutant strain A. Another mutant strain has an insertion site in the third exon of the same specific gene, and this mutant strain is named mutant strain B. Mutants A and B lack the PSI response center protein (PsaA protein), but the levels of PSII core subunit, cytochrome b6f complex, ATP synthase and Rubisco are basically the same as those of the wild plants. The leaves of the plants show increased chlorophyll fluorescence. When the energy of the seed is...

Embodiment 2

[0095] Example 2. Comparison of mutant corn and wild-type corn

[0096] Using the seed embryo of maize HiII as the receptor, the Zm-PSA3 gene was knocked out using CAS9 technology to obtain a mutant maize, which was named Zm-psa3 mutant. After whole-genome sequencing, the Zm-psa3 mutant differs from maize HiII only in that the Zm-PSA3 gene in one chromosome loses its coding function, and the Zm-PSA3 gene in the other chromosome is the same as the receptor.

[0097] The plants to be tested were the homozygous progeny of maize HiII and Zm-psa3 mutants (ie, the Zm-PSA3 genes in both chromosomes have lost their coding function).

[0098] 1. Phenotype identification

[0099] See the phenotype of the plant to be tested at the three-leaf stage figure 1 . figure 1 In the middle, the plant on the left is maize HiII, and the plant on the right is the homozygous progeny of the Zm-psa3 mutant. Compared with maize HiII, the homozygous progeny of Zm-psa3 mutant grows slowly.

[0100] Use mini-PAM...

Embodiment 3

[0105] Example 3 Comparison of wild-type Arabidopsis, mutant Arabidopsis, and complementing strains of Arabidopsis

[0106] The literature mentioning the pCAMBIA1300-flag vector (the corresponding name in the literature is "a modifiedpCAMBIA1300 containing either the Myc or Flag coding sequence"): Zhao Y, Xie S, Li X, Wang C, Chen Z, Lai J, Gong Z. . REPRESSOR OF SILENCING5 Encodes a Member of the Small Heat Shock Protein Family and Is Required for DNA Demethylation in Arabidopsis. Plant Cell. 2014 Jun; 26(6): 2660-2675..

[0107] 1. Obtainment of replenishment strains

[0108] 1. Insert sequence 4 from the 5'end of the double-stranded DNA molecule represented by nucleotides 1-831 from the 5'end between the XbaI and SalI restriction sites of the pCAMBIA1300-flag vector to obtain a recombinant plasmid. According to the sequencing results, the structure of the recombinant plasmid is described as follows: between the XbaI and SalI restriction sites of the pCAMBIA1300-flag vector, the ...

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Abstract

The invention discloses application of PSA3 (photosystem I assembly 3) protein in assembly of PSI (photosystem I) and maintaining of PSI stability. Claimed herein first is the application of calcium homeostasis regulator (PSA3 protein) in: assembly of photosystem I complex (PSI) (a1); formation of the photosystem I complex (PSI) (a2); maintaining of the stability of the photosystem I complex PSI) (a3). It is discovered by the inventor that under the lack of PSA3 protein, the photosystem I complex is unable to be assembled correctly so that the PSA3 protein is degraded fast; in addition, the PSA3 protein and PYG7 protein jointly assists in assembly and stabilization of the PSI complex on matrix side of PSI-assembled thylakoid membrane. The application according to the invention is of great applicable value in the study on photosynthetic eukaryotic PSI.

Description

Technical field [0001] The present invention relates to the application of PSA3 protein in participating in the assembly of PSI and maintaining the stability of PSI. Background technique [0002] In cyanobacteria, algae and plants, there are two types of thylakoid membrane protein complexes, one is the photosystem Ⅰ complex (PSI) and the other is the photosystem II complex (PSII). [0003] PSI is about 600kD and consists of two smaller subcomplexes. Almost half of the subunits of PSI are encoded by the chloroplast genome. The two subcomplexes of PSI are: ①The core complex, which is responsible for transferring electrons from the plastocyanin on the cystic side of the thylakoid to the ferredoxin on the matrix side; ②The LHCI complex, which is responsible for capturing light energy. The core complex consists of 9 membrane proteins (PsaA, PsaB, PsaF-PsaL) and 3 peripheral subunits (PsaC, PsaD and PsaE). The LHCI complex is composed of four Lhca proteins encoded by the nucleus and p...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/01C12N15/82A01H5/00G01N33/68
CPCC07K14/415C12N15/01C12N15/8269G01N33/68G01N33/6845
Inventor 沈杰王柏臣
Owner INST OF BOTANY CHINESE ACAD OF SCI
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