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Lettuce phytol kinase protein coded sequence

An alcohol kinase protein and coding sequence technology, which is applied in the fields of plant gene improvement, botanical equipment and methods, biochemical equipment and methods, etc., can solve problems such as reports about the coding sequence of lettuce phytol kinase protein, etc. The effect of mass production, increasing content and improving nutritional quality

Inactive Publication Date: 2009-08-26
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
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AI Technical Summary

Problems solved by technology

[0007] After searching the literature of the prior art, it is found that there is no report related to the coding sequence of lettuce phytol kinase protein

Method used

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Examples

Experimental program
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Effect test

Embodiment

[0023] Step 1, the cloning of lettuce phytol kinase gene (VTE-5)

[0024] 1. Extraction of RNA

[0025] Lettuce leaf tissue was taken, ground in liquid nitrogen, added to a 1.5mL Eppendorf centrifuge tube filled with lysate, and after full shaking, total RNA was extracted according to the instructions of the TIANGEN kit. The quality of total RNA was identified by formaldehyde denaturing gel electrophoresis, and then the RNA content was determined on a spectrophotometer.

[0026] 2. Full-length cloning of genes

[0027] According to the conserved sequence of amino acids encoded by related genes in Arabidopsis thaliana, the RACE method was used for full-length cDNA cloning, and the Clontech kit was used for cloning in three stages:

[0028] (1) Synthesis of first-strand cDNA

[0029] Using the 5'-CDS primer A and SMART II A oligo primers provided by the Clontech kit, use the extracted total RNA as a template to synthesize 5'-RACE-Ready cDNA under the action of PowerScript rev...

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Abstract

The invention relates to a lettuce phytol kinase protein coded sequence, belonging to the field of genetic engineering technology. The lettuce phytol kinase protein coded sequence has nucleotide sequence shown by 8-937 bits in SEQ ID NO.3, and the polypeptide encoded by the lettuce phytol kinase protein coded sequence has amino acid sequence shown in SEQ ID NO.4; a method for increasing the content of vitamin E by utilizing the lettuce phytol kinase protein coded sequence comprises the steps: the lettuce phytol kinase protein coded sequence is cloned; the cloned sequence is connected on plant expression regulator sequence, so that the corresponding plant expression vector is obtained; the expression vector is transferred agrobacterium to be converted to be arabidopsis; after being screened by antibiotics, transformed cells containing the lettuce phytol kinase protein coded sequence can be obtained; finally, transgene plant or descendants thereof can be regenerated. The invention can increase the content of vitamin E, improve the nutritional quality of foods, ensure the transgene plant containing the phytol kinase protein coded sequence to become a bioreactor, and further realize the mass production of the vitamin E.

Description

technical field [0001] The invention relates to a protein coding sequence in the technical field of genetic engineering, in particular to a lettuce phytol kinase 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 and α-tocotrienol , β-tocotrienol, γ-tocotrienol, and δ-tocotrienol, among which α-tocotrienol has the highest biological activity and is considered to be the main active ingredient of vitamin E. [0003] The biological function of vitamin E was discovered in 1922. Medical research shows that vitamin E is not only related to the reproductive system, but also closely related to the normal metabolism of the central nervous system, digestive system, cardiovascular system and muscular system. Vitamin E is involved in the construction and maintenance of cell membranes, and is an important ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/54C12N15/82
Inventor 唐岳立唐克轩任薇薇
Owner SHANGHAI JIAO TONG UNIV
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