Method for obtaining male and female differential expression genes of early embryos or larvae of prawns

A technique of differentially expressing genes and larvae, which is applied in the fields of molecular biology and genetic breeding, and can solve the problems of distinguishing sex, missing, small size, etc.

Active Publication Date: 2020-04-07
INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, Litopenaeus invana is very small in the early stage of development, sex cannot be distinguished according to phenotype, and RNA cannot be extracted by conventional methods, so no relevant research has been seen so far

Method used

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  • Method for obtaining male and female differential expression genes of early embryos or larvae of prawns
  • Method for obtaining male and female differential expression genes of early embryos or larvae of prawns
  • Method for obtaining male and female differential expression genes of early embryos or larvae of prawns

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Embodiment 1

[0012] Example 1: Establishment of male and female trace transcriptomes and analysis of differentially expressed genes of male and female Litopenaeus vannamei juveniles

[0013] (1) Source of material

[0014] Litopenaeus vannamei juveniles were obtained from Hainan Guangshun Taipu Marine Breeding Co., Ltd., referring to the early developmental morphology manual of Litopenaeus vannamei, and after confirming that they were juveniles under a microscope, single-tailed individuals were picked and placed in 250 μl RNA-free Enzyme centrifuge tubes, and quickly placed in liquid nitrogen. All samples were stored at -80°C for later use.

[0015] (2) DNA and RNA co-extraction

[0016] RNA extraction:

[0017] Add 50 μl Trizol to the centrifuge tube containing the single-tailed individual, after repeated pipetting, add 20 μl chloroform, mix well by inverting up and down, let it stand for 5 minutes, and centrifuge at 12000g at 4°C for 10 minutes;

[0018] Pipette 30-40 μl of the upper...

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Abstract

According to the invention, prawns have obvious sex bimorphism; sex determination and differentiation of the prawns often occur in an early development stage; thus, sex determination and sex differentiation related genes need to be screened in the early development stage of male and female individuals. Since prawn individuals at the early development stage are extremely small, and cannot be distinguished in sex, so a screening method for male and female differential expression genes of early embryos or larvae of the prawns is not established yet. According to the invention, co-extracting of DNA and RNA of a single individual of a prawn zoae is realized; a sex probe is used for sex identification on a DNA level; RNAs of a same gender are mixed to respectively construct a male RNA library and a female RNA library in a zoae stage; a micro transcriptome sequencing technology is utilized to obtain the female and male larva differential expression genes; thus, the screening method of the male and female differential expression genes of the early embryos or larvae of the prawns is established. The screening method provided by the invention promotes the research on sex determination and sex differentiation of the prawns and provides a basis for analyzing a sex determination and differentiation mechanism.

Description

technical field [0001] The invention belongs to the fields of molecular biology and genetic breeding, and in particular relates to a method for obtaining differentially expressed genes of male and female early embryos or juveniles of prawns. Background technique [0002] Gender is one of the important propositions in life sciences. The study of biological sex determination and sex differentiation mechanism will help to enrich the understanding of the evolution process of species sex. At the same time, because many economic animals have significant differences in growth traits between male and female, the study of sex determination and sex differentiation is also of great significance for sex control and parthenogenesis. [0003] Litopenaeus vannamei is the main mariculture species in my country, with an annual output of up to 1.5 million tons. In the mid-to-late growth period of this species, the growth rate of females is significantly faster than that of males. Therefore, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6879C12Q1/6809G16B30/00
CPCC12Q1/6879C12Q1/6809G16B30/00C12Q2600/158C12Q2600/124C12Q2539/113
Inventor 李富花王悦于洋李诗豪
Owner INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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