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Molecular biological method for fish ovarian development quantification by using 5S rRNA (ribosomal ribonucleic acid)

A molecular biology, ovarian development technology, applied in the direction of biochemical equipment and methods, microbial determination/inspection, etc., can solve problems such as time-consuming, long-term training, no quantification of gonadal development, etc., to avoid subjective judgment, quantitative The effect of high chemical degree and protection of precious samples

Active Publication Date: 2022-03-15
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, gonadal histology has three major disadvantages: first, histological procedures are cumbersome and time-consuming (at least 48 hours to complete a batch of samples), including tissue fixation, preservation, dehydration, clearing, wax immersion, embedding, sectioning, patching, More than 20 steps, such as baking and staining, are made into tissue sections, and pictures are taken after observation under a microscope
Second, the identification of various types of oocytes in histology must be carried out by professionals and requires long-term training to complete
Thirdly, for histological tissue, a sufficient amount of gonad tissue is usually obtained after the fish is sacrificed for subsequent processing. Although there are non-lethal methods, the sample size is too small to truly reflect the overall status of ovarian development
However, the GSI at the same gonadal development period may be different among individuals with different weights. GSI is also affected by many environmental factors (such as population density, food abundance, light and water temperature conditions). GSI is also an indicator of allometric growth. There are large differences among individuals in the age group
Therefore, there is currently no method worldwide to quantify gonadal development in fish

Method used

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  • Molecular biological method for fish ovarian development quantification by using 5S rRNA (ribosomal ribonucleic acid)
  • Molecular biological method for fish ovarian development quantification by using 5S rRNA (ribosomal ribonucleic acid)
  • Molecular biological method for fish ovarian development quantification by using 5S rRNA (ribosomal ribonucleic acid)

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Embodiment 1

[0025] Combine below figure 1 , take the fish California perch of oocyte asynchronous development as an example, the present invention is described in further detail:

[0026] A molecular biology method and application thereof for quantitative quantification of California perch ovary development using 5S rRNA, the steps of which are:

[0027] (1) Ovary samples of California perch females at various stages of ovarian development were continuously obtained, including 2-month-old, 3-month-old, 4-month-old, 5-month-old, 6-month-old, 8-month-old, and 10-month-old. Tail fish, take 2 parts of each fish ovary sample in each period, one part takes 50mg and saves it in RNAlater (Sigma China), puts it in the refrigerator at 4℃ overnight, and saves it in the -80℃ refrigerator for later use, and the other part of the ovarian tissue Take 50 mg and fix it in Bouin's solution, transfer it to 70% ethanol after 48 hours, and store it at room temperature for a long time.

[0028] (2) Take 10 m...

Embodiment 2

[0033] Combine below figure 2 , taking the fish golden perch of oocyte synchronous development as an example, the present invention is further described in detail:

[0034] A molecular biology method and its application for quantitative quantification of golden perch ovary development using 5S rRNA, the steps of which are:

[0035] (1) Continuously obtain the ovary samples of golden perch females at various stages of ovarian development, including 2-month-old, 4-month-old, 6-month-old, 8-month-old, and 10-month-old. Take 2 parts of tail fish ovary samples, take 50 mg of one part and store it in RNAlater (Sigma China), put it in the refrigerator at 4 ℃ overnight, and store it in the -80 ℃ refrigerator for later use, and take 50 mg of the other part of ovarian tissue and fix it in Bouin's solution After 48 hours, it was transferred to 70% ethanol, and stored at room temperature for a long time.

[0036] (2) Take 10 mg of each ovary sample stored at -80°C, extract the total RN...

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Abstract

The invention discloses a molecular biological method for fish ovarian development quantification by using 5S rRNA (ribosomal Ribonucleic Acid), which comprises the following steps: acquiring ovarian samples of specific female fish in different development stages, measuring the ratio of 5S rRNA to total RNA in the ovarian samples in each development stage and the ratio of the volume of primary oocytes to the total volume of all oocytes, and constructing a formula of the correlation between the 5S rRNA ratio and the primary oocyte volume ratio, calculating the primary oocyte volume ratio through the 5S rRNA ratio, and judging the female fish ovary development stage according to the primary oocyte volume ratio. The method solves the problems that an effective quantitative method is not established for fish ovarian development at the present stage, ovarian development judgment standards are not clear, subjectivity is high, judgment steps are tedious, consumed time is long, sampling is lethal and the like, and the fish ovarian development stage can be accurately judged by adopting trace ovarian tissues in a non-lethal mode.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a molecular biology method for quantifying fish ovary development by using 5S rRNA. Background technique [0002] my country is a big aquaculture country in the world. Its aquaculture production has ranked first in the world for more than 30 consecutive years, and its aquaculture production accounts for nearly 2 / 3 of the world's aquaculture production. The success of aquaculture in my country is largely due to the efforts of Chinese scientific researchers who have made breakthroughs in artificial breeding techniques for various fish, thus breaking the bottleneck of relying on fishing fry from natural water bodies for breeding. The breakthrough of artificial reproduction technology was realized on the basis of a large number of researches on fish ovary development. The research on gonad development of fish not only contributes to the breakthrough of artificial reproductio...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888
CPCC12Q1/6888C12Q2600/158C12Q2600/178Y02A40/81
Inventor 沈志刚管赫赫于跃刘娅李波丁洋鲁子怡耿龙慧别汉晴胡晨昊
Owner HUAZHONG AGRI UNIV