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Method for obtaining somatic cells by performing enzymolysis on purple laver by using conch digestive glands

A technology for digesting glands and somatic cells, applied in cell dissociation methods, biochemical equipment and methods, plant cells, etc., can solve problems such as high price, achieve long operation cycle, strong disease resistance and stress resistance, and reduce costs Effect

Inactive Publication Date: 2016-06-15
福建省平潭县水产良种实验有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the current enzymatic hydrolysis method requires the use of multiple enzymes such as lyase and cellulase, and most of the enzymes used are imported commodities, which are expensive

Method used

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  • Method for obtaining somatic cells by performing enzymolysis on purple laver by using conch digestive glands

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] 1) Wash fresh corolla snails in seawater for 5 times, starve them in water for 2 days, wash them in seawater for 3 times, crush the conch, take out the digestive glands, and rinse with water for 2 times;

[0024] 2) Put the rinsed digestive gland into a homogenizer for homogenization, and then dissolve it in a phosphate buffer solution with a pH value of 7.0 and a concentration of 0.1mol / L;

[0025] 3) Add ammonium sulfate to the solution obtained in step 2), so that the saturation of ammonium sulfate in the solution is 30%, carry out salting out, dialyze at 4°C overnight, then centrifuge at 1000r / min for 15min, take the supernatant and add ammonium sulfate Carry out secondary salting out until the saturation is 90%, and take the precipitate after dialysis and centrifugation;

[0026] 4) Add water to the precipitate obtained in step 3) to prepare an enzyme solution with a concentration of 0.2 μg / mL;

[0027] 5) Shred laver tissue, add enzyme solution at 1 mL / g, enzymat...

Embodiment 2

[0029] 1) Wash the fresh corolla snails in seawater for 6 times, put them in clean water to starve for 2 days, wash them in seawater for 4 times, smash the conch, take out the digestive glands, and rinse with water for 2 times;

[0030] 2) Put the rinsed digestive gland into a homogenizer for homogenization, and then dissolve it in a phosphate buffer solution with a pH value of 7.0 and a concentration of 0.1mol / L;

[0031] 3) Add ammonium sulfate to the solution obtained in step 2), so that the saturation of ammonium sulfate in the solution is 30%, carry out salting out, dialyze at 4°C overnight, then centrifuge at 1000r / min for 15min, take the supernatant and add ammonium sulfate Carry out secondary salting out until the saturation is 90%, and take the precipitate after dialysis and centrifugation;

[0032] 4) Add water to the precipitate obtained in step 3) to prepare an enzyme solution with a concentration of 0.2 μg / mL;

[0033] 5) Shred laver tissue, add enzyme solution a...

Embodiment 3

[0035] 1) After washing the fresh Korean corolla snails in seawater for 8 times, put them in clean water to starve for 2 days, wash them in seawater for 5 times, smash the conch, take out the digestive glands, and rinse them with water for 2 times;

[0036] 2) Put the rinsed digestive gland into a homogenizer for homogenization, and then dissolve it in a phosphate buffer solution with a pH value of 7.0 and a concentration of 0.1mol / L;

[0037] 3) Add ammonium sulfate to the solution obtained in step 2), so that the saturation of ammonium sulfate in the solution is 30%, carry out salting out, dialyze at 4°C overnight, then centrifuge at 1000r / min for 15min, take the supernatant and add ammonium sulfate Carry out secondary salting out until the saturation is 90%, and take the precipitate after dialysis and centrifugation;

[0038] 4) Add water to the precipitate obtained in step 3) to prepare an enzyme solution with a concentration of 0.3 μg / mL;

[0039] 5) Shred laver tissue, ...

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Abstract

The invention discloses a method for obtaining somatic cells by performing enzymolysis on purple laver by using conch digestive glands. The method comprises the following steps: homogenizing fresh conch digestive glands, carrying out salting-out, and carrying out enzymolysis on purple laver tissues by directly using the salting-out product as the enzyme solution, thereby obtaining the somatic cells. The treatment method can simplify the operation and shorten the enzymolysis time, has favorable enzymolysis effect, does not need to add cellulase or other enzymes in the enzymolysis process, can effectively lower the cost, has favorable economic benefits, and is suitable for industrialized production and popularization.

Description

technical field [0001] The invention belongs to the technical field of marine product cultivation, and in particular relates to a method for obtaining somatic cells by enzymolyzing seaweed with conch digestive glands. Background technique [0002] Seaweed is the general name of alternate algae in the sea. It is rich in protein, iodine, phosphorus, calcium and other nutrients. It is also a kind of cultivated seaweed with the highest economic value in the world at present, and its output value accounts for about two-thirds of the total output value of artificially cultivated seaweed in the world. [0003] Select mature laver algae with excellent characteristics from natural populations or cultivated populations every year as seed algae, and then obtain fruit spores from the seed algae or obtain somatic cells by rotting method, block method or enzymatic hydrolysis method, and then cultivate them into filaments. Inoculating shells and cultivating shell filaments in large quanti...

Claims

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Application Information

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IPC IPC(8): C12N5/04A01G33/02
CPCC12N5/04A01G33/00C12N2509/00
Inventor 高丽华高文刘燕飞
Owner 福建省平潭县水产良种实验有限公司
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