Method for analyzing hypoxia stress on shrimps by detecting HURP1 gene expression

A gene expression, shrimp technology, applied in the field of molecular biology, can solve the problem of determining the hypoxic stress, shrimp farming loss, growth rate, and adverse effects of immune function in Littoria vannamei

Inactive Publication Date: 2017-04-26
SUN YAT SEN UNIV
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Problems solved by technology

Insufficient dissolved oxygen can easily cause hypoxic stress to Litopenaeus vannamei, which has adverse effects on the growth rate and immune function of Litopenaeus vannamei, and often causes losses to the shrimp farming industry
Since the absorption and utilization of dissolved oxygen in water by prawns is affected by many other physical and chemical factors, such as temperature or ammonia nitrogen concentratio

Method used

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  • Method for analyzing hypoxia stress on shrimps by detecting HURP1 gene expression
  • Method for analyzing hypoxia stress on shrimps by detecting HURP1 gene expression
  • Method for analyzing hypoxia stress on shrimps by detecting HURP1 gene expression

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Embodiment

[0024] 1. Acquisition of experimental samples

[0025] Select 400 Litopenaeus vannamei (about 7g / head) with uniform specifications, and randomly divide them into experimental group and control group equally, with 200 prawns in each group. The control group was fully oxygenated and tested every two hours with a dissolved oxygen meter to ensure that the dissolved oxygen in the water was above 5mg / L; the experimental group used the method of filling the water with nitrogen combined with oxygen to control the dissolved oxygen in the water to 1.2mg / L (generally considered Dissolved oxygen in water is 1.2 mg / L (the lower limit of dissolved oxygen acceptable to Litopenaeus vannamei), which is tested by a dissolved oxygen meter every two hours. When the dissolved oxygen in the water of the experimental group decreased to 1.2 mg / L, the timing began, and the hemolymph samples of Litopenaeus vannamei were taken at 3, 6, 9, 12, 18, 24, 36, 48 and 72 hours after the experiment (every 5 shr...

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Abstract

The invention provides a method for analyzing hypoxia stress on shrimps by detecting HURP1 gene expression. The method comprises the steps as follows: step one, acquiring the sequence of an open reading frame of an HURP1 gene of litopenaeus vannamei, and designing and synthesizing a sequence-specific fluorescent quantitative PCR (polymerase chain reaction) primer; step two, acquiring to-be-detected blood lymphocytes of litopenaeus vannamei, extracting the total RNA (ribose nucleic acid) of the blood lymphocytes, reversely transcribing the total RNA into cDNA, and preparing a fluorescent quantitative PCR template; step three, performing fluorescent quantitative PCR analysis to analyze expression of HURP1 in the blood lymphocytes of litopenaeus vannamei. With the adoption of the method, whether litopenaeus vannamei is in a low-oxygen stress state is judged more conveniently, more quickly and more accurately by detecting expression of the HURP1 gene.

Description

technical field [0001] The invention belongs to the field of molecular biology, and in particular relates to a method for analyzing whether Litopenaeus vannamei is in a state of hypoxic stress by detecting the expression of HURP1 gene. Background technique [0002] The prawn (Litopenaeus vannamei) in the genus Penaeus is an important species of aquaculture in my country. In 2015, the global production of farmed prawns reached 3.3 million tons, of which, the output of farmed prawns in my country was 1.5 million tons, making it the largest country in the world for prawn farming. my country mainly cultures four species of prawns, Litopenaeus vannamei, Penaeus monodon, Fenneropenaeus chinensis and Marsupenaeus japonicus, among which Litopenaeus vannamei is the main cultured species, about It accounts for 90% of the total production of cultured prawns. [0003] In shrimp culture water, oxygen is consumed by various biological or abiotic factors, and the proportion of oxygen cons...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6851C12Q1/6888C12Q2600/158
Inventor 陈义烘翁少萍何建国
Owner SUN YAT SEN UNIV
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