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Lettuce tocopherol cyclase protein coding sequence

A technology of phenol cyclase protein and coding sequence, which is applied in the fields of biochemical equipment and methods, lyase, application, etc., can solve the problems that have not yet been reported about the coding sequence of lettuce tocopherol cyclase protein, and achieve the goal of increasing the content of tocopherol cyclase , the effect of improving nutritional quality

Inactive Publication Date: 2011-06-08
SHANGHAI JIAOTONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] After searching the literature of the prior art, it is found that there is no report related to the protein coding sequence of lettuce tocopherol cyclase

Method used

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Experimental program
Comparison scheme
Effect test

Embodiment

[0020] Step 1, cloning of lettuce tocopherol cyclase gene

[0021] (1) Extraction of RNA

[0022] Lettuce leaf tissue was taken, ground in liquid nitrogen, added to a 1.5mL Eppendorf (EP) centrifuge tube filled with lysate, and fully oscillated, total RNA was extracted according to the instructions of the TIANGEN kit, and the total RNA was identified by formaldehyde denaturing gel electrophoresis. RNA quality was then measured on a spectrophotometer for RNA content.

[0023] (2) Full-length cloning of the gene

[0024] According to the conserved sequence of amino acids encoded by related genes in Arabidopsis, using the principle of homologous gene cloning, the RACE (Rapid Amplification of cDNA Ends) method was used to clone the full-length cDNA. Cloning was performed using Clontech kits, and the cloning was carried out in three stages. :

[0025] ①Synthesis of first-strand cDNA

[0026] Using the 5'-CDS primer A and SMART II A oligo primers provided by the Clontech kit, us...

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PUM

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Abstract

The invention provides a lettuce tocopherol cyclase protein-coding sequence in the technical field of genetic engineering, which has a nucleotide sequence shown in a 6-1526th position in SEQ ID NO.3, as well as a degenerate sequence thereof. Code of the coding sequence has an amino acid sequence shown in SEQ ID NO.4, conservative mutation polypeptide, active fragments or active derivatives thereof. A method for increasing the VE content of plants by utilizing the coding sequence comprises the following steps: connecting the lettuce tocopherol cyclase protein-coding sequence to a plant expression regulatory sequence so as to obtain a plant expression vector containing the lettuce tocopherol cyclase protein-coding sequence; transferring the expression vector into agrobacterium; utilizing the agrobacterium to transform Arabidopsis; obtaining a transformed cell containing the lettuce tocopherol cyclase protein-coding sequence by screening antibiotics; and regenerating a transgenic plant and progeny thereof. The invention has the advantages of increasing the VE content of vegetables and realizing the large-scale production of VE.

Description

technical field [0001] The invention relates to a protein coding sequence in the technical field of genetic engineering, in particular to a lettuce tocopherol cyclase protein coding sequence. Background technique [0002] Vitamin E is a fat-soluble vitamin, and its natural products are divided into eight types according to the structure, namely α-tocopherol, β-tocopherol, γ-tocopherol, δ-tocopherol (tocopherol) and α, β , γ, δ-tocotrienol (tocotrienol), of which α-tocopherol has the highest biological activity and is considered to be the main active ingredient of vitamin E. Vitamin E has an important impact on improving the body's immunity. It has anti-aging functions, can eliminate cytochrome deposition, improve skin elasticity, and weaken gonadal atrophy. Therefore, it is widely used in a large number of health care products and beauty products. Vitamin E can prevent and treat a variety of gynecological diseases, treat hemolytic anemia and favism in premature infants, and...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/60C12N9/88C12N15/82
Inventor 唐岳立唐克轩任薇薇王玥月
Owner SHANGHAI JIAOTONG UNIV