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Phosphoenolpyruvate carboxylase gene of watergrass and its coded protein and uses

A technology of pyruvate carboxylase and phosphoenol, which is applied in the directions of enzymes, biochemical equipment and methods, and the introduction of foreign genetic material using carriers to achieve the effect of improving the efficiency of photosynthesis

Inactive Publication Date: 2006-11-22
INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Research on the development and utilization of barnyardgrass resources and the cloning of photosynthetic functional genes is still blank in the world

Method used

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  • Phosphoenolpyruvate carboxylase gene of watergrass and its coded protein and uses
  • Phosphoenolpyruvate carboxylase gene of watergrass and its coded protein and uses
  • Phosphoenolpyruvate carboxylase gene of watergrass and its coded protein and uses

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Embodiment 1, the cloning of barnyardgrass phosphoenolpyruvate carboxylase gene Eppc

[0044] 1. Extraction of barnyardgrass total RNA and synthesis of eDNA

[0045] Take the green leaves of barnyardgrass plants after 4 hours of light, use the Trizol kit (Gibeco company) and refer to the kit instructions to extract total RNA, the specific method is: cut 1g of fresh barnyardgrass leaves, put them in liquid nitrogen and grind them quickly to freeze-dry Put it into a 1.5mL centrifuge tube and add 1mL Trizol reagent, mix well, leave at room temperature for 5min, centrifuge at 12000g for 10min at 4°C, transfer the supernatant to another centrifuge tube, place at room temperature for 5min, then add 0.2mL chloroform, Shake for 15 sec, place at room temperature for 5 min, centrifuge at 12,000 g at 4°C for 15 min, transfer the supernatant to another centrifuge tube, add 0.5 mL isopropanol, place at room temperature for 10 min, centrifuge at 12,000 g at 4°C for 10 min, discard th...

Embodiment 2

[0051] Example 2. Functional verification of barnyardgrass phosphoenolpyruvate carboxylase gene Eppc

[0052] One, the construction of the plant expression vector containing Eppc

[0053] 1. Construction of plant expression vector pUbi-Eppc

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Abstract

The invention discloses a barnyard grass phosphoenolpyruvate carboxylase gene and coded protein and appliance, which is one ribotide sequence in the following sequences: 1) sequent graph SEQ ID No.:1;2) DNA sequence hybrid ribotide sequence limited by SEQ ID No.:1. the coded gene protein is one of SEQ ID No.:2 in the 1) sequence graph; 2) SEQ ID No.:2 in the 2) sequence graph with two amino residue sequence replaced, lost or added form one to ten to improve plant photosynthesis efficiency.

Description

technical field [0001] The invention relates to a plant gene and its coded protein and its application, in particular to a phosphoenol pyruvate carboxylase gene and its coded protein and its application in improving plant photosynthesis efficiency. Background technique [0002] C in photosynthetic carbon assimilation in plant kingdom 3 pathway and C 4 Path distinction. Due to the existence of photorespiration, C 3 A considerable part of the carbon fixed by the pathway is consumed by photorespiration, and the formation of biomass is reduced by more than 50%. C 4 Fixing CO in the photosynthetic pathway 2 phosphoenolpyruvate carboxylase (PEPCase) for CO 2 affinity far exceeds that of C 3 Ribulose bisphosphate carboxylase (RuBPase) in the photosynthetic pathway, and phosphoenolpyruvate carboxylase has the ability to concentrate CO 2 mechanism, CO 2 The increase of the concentration is beneficial to the carboxylation activity of the bifunctional enzyme RuBPase and inhibi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/52C12N9/00C12N15/63C12N15/82
Inventor 赵明张桂芳丁在松
Owner INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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