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Multi-stress-induced promoter derived from alpine ash and application of promoter

A promoter and inducible technology, applied in the field of genetic engineering, can solve problems such as inhibiting plant growth and development, and achieve the effect of saving money and improving the ability to resist various adversities

Inactive Publication Date: 2014-02-19
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although these promoters can enable the high-efficiency expression of foreign genes in plants, although the stress resistance of transgenic plants has been improved, they often inhibit the growth and development of plants.

Method used

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  • Multi-stress-induced promoter derived from alpine ash and application of promoter
  • Multi-stress-induced promoter derived from alpine ash and application of promoter
  • Multi-stress-induced promoter derived from alpine ash and application of promoter

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] promoter discovery

[0033] Genomic DNA of Eucalyptus grandis (variety EG6) was extracted, and the extraction of Eucalyptus genomic DNA referred to conventional methods in the art. Design specific primers (P1: 5'-TCAT TCTAGA GCGTAATTGCTGTC GCACCC-3', such as SEQ ID NO: 2, the underline is the Xba I restriction site) and (P2: 5'-GCTC AAGCTTCGAGAAGGAATCGCTTCTGA-3', such as SEQ ID NO: 3, the underline is the Sac I restriction site) to amplify the genomic DNA, recover and clone the fragments, and perform DNA sequencing to obtain a 720bp DNA fragment, the 720bp DNA fragment The nucleotide sequence is shown in SEQ ID NO:1. The amplified sequence shown in SEQ ID NO: 1 was analyzed for the regulatory region, the start codon ATG of the gene was identified, and the part of the structural gene was removed. Obtain the regulatory sequence at the 5' end of the gene, the regulatory elements of the promoter such as figure 2 As shown, the TATA box is located in the ?150-?155 regi...

Embodiment 2

[0035] 1. Low temperature and drought-induced activity verification of the promoter pG5

[0036] Construction of recombinant expression vector

[0037] 1. Digest pGEM-T: pG5 prepared in Example 1 with restriction endonucleases Xba I and Sac I, and recover the digested product (pG5).

[0038] 2. Digest the binary vector pBI 121 with restriction endonucleases Xba I and Sac I to obtain the pBI 121 vector backbone with the CaMV 35S promoter removed.

[0039] 3. Ligate the digested product of step 1 with the vector backbone of step 2 to obtain a recombinant plasmid, and perform sequencing verification. The sequencing results show that the target plasmid pG5-GUS (replacing pBI with pG5 of the sequence shown in SEQ ID NO: 1) The small fragment between the recognition sites of Xba I and Sac I of 121).

[0040] 2. Transformation of tobacco by transient expression

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Abstract

The invention belongs to the technical field of genetic engineering and particularly discloses a multi-stress-induced promoter derived from alpine ash and an application of the promoter. The promoter is specifically a promoter of a multi-stress-induced gene G6PDH of an alpine ash variety EG6, and a nucleotide sequence of the promoter is as shown in SEQ ID NO: 1. A regulation and control element of the promoter consists of a low-temperature stress response element, a draught stress response element, an optical response element, a hormone response element and other elements. The promoter can be subjected to induced expression by stresses, such as low temperature and draught, can be used for botanical induced expression vectors and has wide applicability and good application prospect in the field of bioengineering.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to a multi-stress-inducible promoter pG5 derived from Eucalyptus macrophylla and its application. Background technique [0002] The pentose phosphate pathway is an important pathway for sugar metabolism in plants. Its main physiological function is to provide reducing power NADPH for biosynthesis and five-carbon sugar for nucleic acid synthesis, and some intermediate products can participate in the synthesis of amino acids and fatty acids, etc. Glucose-6-phosphate dehydrogenase is its key regulatory rate-limiting enzyme (Lin Yuanzhen. 2006. Gene cloning, structural analysis and functional identification of sweet poplar glucose-6-phosphate dehydrogenase. Doctoral dissertation, Beijing Forestry University , Supervisor: Zhang Zhiyi, pp.44-66), involved in the plant response caused by various environmental stresses, such as the interaction with some metal ions (such as Al 3...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C12N15/63C12N1/15C12N1/19C12N1/21C12N5/10A01H5/00
Inventor 林元震莫晓勇杨小红杨会肖周玮陈晓阳
Owner SOUTH CHINA AGRI UNIV