Method of large yellow crocker sperm genetic substance complete deactivation

A technology for genetic material and large yellow croaker, which is applied in the fields of biochemical equipment and methods, microorganisms, and treatment of microorganisms with electricity/wave energy, etc. simple effect

Inactive Publication Date: 2007-07-04
INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Applying the genetic inactivation conditions of flounder and flounder sperm directly to the genetic inactivation of large yellow croaker sperm has not been successful, and its genetic inactivation rate is only 88-92%. The method experiment report has not yet been seen

Method used

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  • Method of large yellow crocker sperm genetic substance complete deactivation
  • Method of large yellow crocker sperm genetic substance complete deactivation
  • Method of large yellow crocker sperm genetic substance complete deactivation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] The inactivation steps are:

[0026] 1. Semen preservation: artificially extruded mature semen of inactivated large yellow croaker was stored in the dark at 2°C for later use.

[0027] 2. Semen dilution: Dilute the mature semen of large yellow croaker with a pre-cooled diluent, and the dilution factor is 40 times, and then put the diluted mature semen of large yellow croaker into a petri dish that has been pre-cooled to 0°C. The liquid thickness of the mature semen of large yellow croaker diluted in the petri dish is 0.1 cm.

[0028] Wherein the pre-cooled diluent is NaCl 0.60% by weight volume percentage; KCl 0.03%; CaCl 2 0.01%; Glucose 0.05%; pH7.0; The diluent is pre-cooled to 0°C.

[0029] 3. Inactivation of sperm genetic material: After turning on the ultraviolet light source and preheating for 5 minutes, place the petri dish with diluted large yellow croaker mature semen under the ultraviolet light source, keep the distance between the ultraviolet light source...

Embodiment 2

[0032] The difference from Example 1 is:

[0033] 1. Semen preservation: artificially extruded mature semen of inactivated large yellow croaker was stored in the dark at 3°C ​​for later use.

[0034] 2. Semen dilution: Dilute the mature semen of large yellow croaker with a pre-cooled diluent, and the dilution factor is 50 times, and then put the diluted mature semen of large yellow croaker into a petri dish that has been pre-cooled to 1°C. The liquid thickness of the mature semen of large yellow croaker after dilution in the petri dish is 0.2cm;

[0035] The pre-cooled diluent is NaCl 0.75%, KCl 0.04%, CaCl 2 0.02%, glucose 0.08%; pH7.2; diluent pre-cooled to 1 ℃.

[0036] 3. Inactivation of sperm genetic material: After turning on the ultraviolet light source and preheating for 8 minutes, place the petri dish with diluted large yellow croaker mature semen under the ultraviolet light source, keep the distance between the ultraviolet light source and the bottom of the petri ...

Embodiment 3

[0039] The difference from Example 1 is:

[0040] 1. Preservation of semen: The artificially extruded mature semen of large yellow croaker without activation was stored in the dark at 4°C for later use.

[0041] 2. Semen dilution: Dilute the mature semen of large yellow croaker with a pre-cooled diluent, and the dilution factor is 80 times, and then put the diluted mature semen of large yellow croaker into a petri dish that has been pre-cooled to 2°C. The liquid thickness of the mature semen of large yellow croaker after dilution in the petri dish is 0.25cm;

[0042] The pre-cooled diluent is based on weight volume percentage, NaCl 0.80%, KCl 0.05%, CaCl 2 0.03%, glucose 0.10%; pH 7.3; the diluent is pre-cooled to 2°C.

[0043] 3. Inactivation of sperm genetic material: After turning on the ultraviolet light source and preheating for 10 minutes, place the petri dish with the diluted large yellow croaker mature semen under the ultraviolet light source, keep the distance betw...

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Abstract

The invention discloses a method for complete inactivating germ genetic substance for yellow croaker, which can be used for research and production foryellow croaker gynogenesis and whole femal seed induction. The method comprises following steps: diluting the mature sperm of yellow croaker for 2- 100 times with precooled nutriment- containing diluting liquid; putting it on bottom culturing utensil; irradiating with violet light and impregnating with spawn and incubating; then identifying the effect of sperm genetic inactivation. The invention is characterized by simple method, convenient operation, and 100% of the inactivating rate for sperm genetic substance.

Description

technical field [0001] The invention relates to a technique for manipulating the chromosomes of seawater fish, in particular to a method for completely inactivating the genetic material of large yellow croaker sperm, an important seawater culture object in my country. Background technique [0002] The complete inactivation of sperm genetic material is the key technology to produce homogeneous and heterogeneous gynogenetic diploids in combination with the second polar body of fertilized eggs and the first cleavage inhibition technology. Although there is already a method for ultraviolet genetic inactivation of seawater fish sperm, it is mainly for flounder, and the characteristics of the sperm of large yellow croaker are completely different from those of flounder. The microstructure is quite different from the latter, which makes the sensitivity and tolerance of large yellow croaker sperm to ultraviolet radiation also very different. Applying the genetic inactivation condit...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/06C12N15/01C12N13/00
Inventor 许建和尤锋张培军吴雄飞徐永立蒋宏雷
Owner INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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