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Early light-inductive protein gene and preparation method thereof

A light-induced, protein technology, applied in the fields of botany equipment and methods, biochemical equipment and methods, genetic engineering, etc., can solve the problem of low salt tolerance, and achieve the effect of enhancing salt tolerance and improving salt tolerance.

Inactive Publication Date: 2009-02-11
NORTHEAST FORESTRY UNIVERSITY
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  • Summary
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  • Claims
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AI Technical Summary

Problems solved by technology

[0003] The purpose of the present invention is to solve the problem that the salt-tolerant transgenic plants obtained through the transformation of existing salt-tolerant genes have low salt-tolerant ability, and provide a kind of early light-induced protein (ELIP) gene and its preparation method

Method used

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  • Early light-inductive protein gene and preparation method thereof
  • Early light-inductive protein gene and preparation method thereof
  • Early light-inductive protein gene and preparation method thereof

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specific Embodiment approach 1

[0017] Embodiment 1: The sequence of the coding region of the early light-inducible protein gene in this embodiment is shown in SEQ ID NO:1.

specific Embodiment approach 2

[0018] Specific embodiment 2: The amino acid sequence of the early light-inducible protein gene in this embodiment is shown in SEQ ID NO: 2.

specific Embodiment approach 3

[0019] Specific embodiment three: In this embodiment, the early light-induced protein gene is prepared according to the following steps:

[0020] 1. The total RNA of Tamarix tamarix was extracted by CTAB method;

[0021] 2. Isolate mRNA with the mRNA isolation kit of Promega Company, and take 5ug of mRNA for constructing Tamarix cDNA library. The cDNA library construction kits are ZAP-cDNA Synthesis Kit and ZAP-cDNA GigapackIII Gold Cloning Kit. 7.2×10 5 pfu, the recombination rate is 98%, and the length of the library clone insert is 0.9-1.1kb;

[0022] 3. Sequence the library clone: ​​the sequencing primer is T3 primer, and the sequencer for cDNA library cloning is MegaBACE from GE TM 1000 DNA sequence analyzer, through the Blastx analysis of library clones to obtain the 5' fragment of the early light-induced protein gene, the sequence is:

[0023] CAGAATTTTAAAATTTTCTTTTTTTATTTATCTATTTATTTCATTGCAAAATTTGCTATACACTAGAATATTCACTAATATTGCATTGTGCACATTCAGCGTAACAAATTTTGTAAATCAAAGC...

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Abstract

An early stage photoinduction protein gene and a preparation method relate to a protein gene and a preparation method, and solve the problem that the salt tolerance of salt tolerant transgenic plant obtained by the existing salt tolerant gene transformation is poor. The coded area sequence of the early stage photoinduction protein gene of the invention is shown as SEQ ID NO: 1. The amino acid sequence of the early stage photoinduction protein gene of the invention is shown as SEQ ID NO: 2. The invention adopts cDNA library technology and suppression subtractive hybridization technique, and carries out the EST analysis on the library clone; 5 and 3' part sequences of the gene are obtained by screening, and then the early stage photoinduction protein gene is obtained by splicing the 5 and 3' sequences. When the gene obtained by the invention is shifted to model plant tobacco, transgene tobacco can regularly grow on a culture medium containing 0.6% of NaCl, the grow situation and physiological index is obviously good compared with the contrast, and the salt tolerance is improved by over 50%.

Description

technical field [0001] The invention relates to a protein gene and a preparation method thereof. Background technique [0002] The rapid development of molecular biology technology provides a powerful technical means for cultivating plants with strong saline-alkali tolerance. At present, scholars at home and abroad have studied the damage of salt to plants and the mechanism of plant salt tolerance, cloned some salt-tolerant genes, and obtained some salt-tolerant transgenic plants through the transformation of these salt-tolerant genes, but the salt-tolerant ability is not high. , affecting the practical application, it can be seen that whether an effective salt-tolerant gene can be cloned is still the key to salt-tolerant genetic engineering breeding. Contents of the invention [0003] The purpose of the present invention is to solve the problem that the salt-tolerant transgenic plants obtained by transformation of the existing salt-tolerant gene have low salt-tolerant ab...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C12N15/10C07K14/415
Inventor 高彩球王玉成姜静刘桂丰杨传平
Owner NORTHEAST FORESTRY UNIVERSITY
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