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Divinyl reductase from corn and coding gene and application thereof

A technology of diethylene and reductase, applied in the direction of oxidoreductase, application, genetic engineering, etc., can solve the problems of not getting

Inactive Publication Date: 2013-07-31
SICHUAN AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, two kinds of diethylene (DV type) substances, diethylene Mg-protoporphyrin IX (DV Mg-proto) and diethylene Mg-protoporphyrin IX monomethyl ester (DV MPE), are converted into the corresponding monoethylene (MV type ) substances have not been confirmed by in vitro enzymatic experiments using purified DVR enzyme proteins, and it is unclear whether these divinyl intermediates are catalyzed by a broadly specific divinyl reductase or The corresponding monoethylene species are catalyzed by multiple discinyl reductases (each with a specific substrate)

Method used

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  • Divinyl reductase from corn and coding gene and application thereof
  • Divinyl reductase from corn and coding gene and application thereof
  • Divinyl reductase from corn and coding gene and application thereof

Examples

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Embodiment 1

[0031] Example 1. Cloning of maize ethylene reductase gene and functional verification of its encoded protein

[0032] 1. Cloning of maize ethylene reductase gene

[0033] Design the following primers:

[0034] ZmDVR-F: 5'-CC GGATCC ATGGCGACCATCCTCTCTATC-3' (the underline is the BamH Ⅰ restriction site)

[0035] ZmDVR-R: 5'-ATG GAATTC GCCTAGAAGATGGTCTGCTC-3' (the underline is the EcoR Ⅰ restriction site)

[0036] Using the DNA extracted from the leaves of the corn (Zea mays L.) variety "B73" inbred line as a template, PCR amplification was performed with primers ZmDVR-F and ZmDVR-R to obtain a 1206bp fragment, which was ligated into pMD-18- The pMD-ZmDVR was constructed on the T(TaKaRa) vector, and then transformed into Escherichia coli JM109 strain, and the positive clones were screened and then sequenced. Sequencing results showed that the amplified fragment had the sequence shown in Sequence 1 in the Sequence Listing, and it was named ZmDVR. The sequence shown in se...

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Abstract

The invention discloses a divinyl reductase (DVR) from corn and coding gene and application thereof. The DVR provided by the invention is a protein shown as 1) or 2): 1) a protein comprising an amino acid sequence shown as a sequence 2 in the sequence table; and 2) a protein derived from 1), with DVR activity and obtained by substituting and / or deleting and / or adding one or more amino acid residues on the sequence 2 in the sequence table. Experiments have proved that the DVR provided by the invention can converse DV chlorophyll a into MV chlorophyll a, converse DV chlide a into MV chlide a, converse DV Pchlide a into MV Pchlide a, converse DV MPE into MV MPE and converse DV Mg-proto into MV Mg-proto.

Description

technical field [0001] The invention relates to a diethylene reductase derived from corn, its coding gene and application. Background technique [0002] Chlorophyll is an important pigment for photosynthesis in plants. Its function is to capture light energy and transfer light energy to the reaction center, so it plays an important role in the growth and yield of plants. According to the number of ethylene side chains, chlorophyll can be divided into 3,8-divinyl chlorophyll (DV Chls) and 3-vinyl chlorophyll (monoethylene chlorophyll, MV Chls). Under normal circumstances, the chlorophyll used by almost all plants and bacteria for photosynthesis is MV chls, and only one type of marine organisms, namely “the marine prochlorophyte Prochlorococcus marinus”, has been found to use DVChls (Chisholm SW, Frankel SL, Goericke R , Olson RJ, Palenik B, Waterbury JB, West-Johnsrud L, Zettler ER (1992) Prochlorococcus marinus nov.gen.nov.sp.: A marine prokaryote containing divinylchloroph...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/02C12N15/53C12N15/63C12N5/10C12N1/15C12N1/19C12N1/21C12P17/18
Inventor 王平荣邓晓建万春美孙昌辉王萍渝马晓智肖云华徐正君朱建清高晓玲
Owner SICHUAN AGRI UNIV