Method for producing transformed plant cells containing recombinant human alkaline phosphatase and using transformed plant cells containing recombinant human alkaline phosphatase
A technology for transforming plants and phosphatases, which can be applied to plant cells, botanical equipment and methods, biochemical equipment and methods, etc., and can solve problems such as unusable food products or food ingredients, restrictions, etc.
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Embodiment 1
[0074] Example 1 Production of carrot food based on human AP.
[0075] In order to introduce the human intestinal AP gene into the plant expression vector, the nucleotide sequence of the human AP recombinant gene with a size of 1587 base pairs that is completely consistent with the nucleotide sequence of the natural mRNA of the human alkaline phosphatase gene was generated by synthesis acid sequence. A restriction site Bgl II sequence (agatct) was then added to the sequence of the recombinant AP gene at the 5' end before the translation initiation site (atg) and a stop codon (tga) at the 3' end The sequence with the restriction site Xba I (atctagaat) was added later. The resulting nucleotide sequence of 1602 base pairs was cloned into the plasmid pAL-T vector. A nucleotide sequence of 1600 base pairs in size was excised from the restriction sites Bgl II and Xba I of the generated pAL-T-AP plasmid and ligated into the earlier generated plasmid p35S-NLS-recA-licBM3 , result...
Embodiment 2
[0082] Example 2. Production of stevia food products based on human AP.
[0083] Plant foods based on recombinant human AP were produced as described in Example 1, but using Stevia rebaudiana as the production plant and somatic embryogenesis to grow AP-containing cells. For the production of Stevia somatic embryos, cell suspensions were cultured in liquid nutrient medium MSM containing 0.2 mg / L indole-3-acetic acid and kinetin. Then, to obtain a homogeneous population of somatic embryos, the suspension culture was filtered through a nylon mesh with a mesh size of 120 mcm, and then through a nylon mesh with a mesh size of 50 mcm. Transfer the cell pellet remaining on the second sieve to fresh medium to form embryos. On average up to 70,000 embryos can be generated from 1 L of suspension.
[0084] It can be used to restore the damaged immune system by adding the obtained food to food or drink heated at not more than 40°C, and the method is 0.5 to 1 bag per person per day.
Embodiment 3
[0085] Example 3. Production of lettuce based foods containing human AP.
[0086] Plant food based on recombinant human AP was produced as described in Example 1, but lettuce (Latucasativa) was used as the production plant and the GV3101 p35S-AP strain was used as the Agrobacterium strain to clone the human secreted AP gene into a plant expression vector, This Agrobacterium strain was generated by using E. coli cells with p35S-AP plasmid as donors, E. coli cells of HB101 pRK2013 strain as promoters of conjugative transfer, and Agrobacterium tumefaciens cells of GV3101 strain as recipients.
[0087] The produced food is used to alleviate the consequences of poisoning, including poisoning related to food poisoning, intestinal infection or drug use, by taking 1-2 bags orally per person per day for 7-10 days.
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