A kind of macrobrachium rosenbergii estrogen-related receptor gene err and its application

A technology of Macrobrachium rosenbergii and receptor genes, applied in the field of genetic engineering, can solve problems such as unfavorable widespread use and complicated operation of yeast two-hybrid system, and achieve the effects of low cost, convenient source, and increased gene expression

Active Publication Date: 2019-10-25
海南永贺生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the operation of the yeast two-hybrid system is relatively complicated, which is not conducive to widespread use

Method used

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  • A kind of macrobrachium rosenbergii estrogen-related receptor gene err and its application
  • A kind of macrobrachium rosenbergii estrogen-related receptor gene err and its application
  • A kind of macrobrachium rosenbergii estrogen-related receptor gene err and its application

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] 1. Extraction of total RNA from Macrobrachium rosenbergii

[0035] Healthy Macrobrachium rosenbergii was taken, anesthetized in an ice bath for about 2 minutes, dissected and sampled, and the ovarian tissue was immediately separated. The total RNA of Macrobrachium rosenbergii was extracted by Trizol reagent method, and its OD 260 / 280 =1.91, electrophoresis result ( figure 1 ) shows that 28S rRNA and 18S rRNA bands are clear, and the brightness of 28S band is twice that of 18S, indicating that the obtained total RNA is not contaminated by protein, phenol and genomic DNA, and has high purity.

[0036] 2. Synthesis of the first strand of cDNA

[0037] 1) Take out 2 μg of preserved Macrobrachium rosenbergii RNA, add 1 μL of polythymine Oligo(dT) (10 mM), mix well, and place in a water bath at 70°C for 5 minutes;

[0038] 2) After standing on ice for 5 minutes, add 2 μL of dNTP Mixture (10 mM), 4 μL of 5XRTBuffer, 1 μL of RNase inhibitor (10 U / μL), and RNase-free ddH to t...

Embodiment 2

[0059] Tissue expression of the M. rosenbergii ERR gene.

[0060] Semi-quantitative PCR was used to detect the expression of the ERR gene of Macrobrachium rosenbergii in various tissues, and the results were as follows: Figure 6 As shown in , the results showed that ERR mRNA was highly expressed in the heart, brain, and ovary; the expression was weak in the hepatopancreas, testis, and gill; no expression signal was detected in the muscle and eye stalk. It shows that ERR participates in regulating the expression of neural regulation and other endocrine regulation genes in Macrobrachium rosenbergii, which enriches the system signaling pathway theory of shrimp endocrine regulation, and can provide a certain theoretical basis for the study of related shrimp sex regulation.

Embodiment 3

[0062] Expression changes of ERR gene and protein in Macrobrachium rosenbergii during NP bathing.

[0063] Allocate healthy female Macrobrachium rosenbergii at the early stage of sexual differentiation into 4 independent plastic buckets (50L) of the same size. After one week of domestication, add NP (nonylphenol) so that the final concentrations are 0, 5, 25, and 125 μg respectively. / L. Feed commercial feeds at 07:00 and 17:00 every day, keep the water temperature, light and other conditions the same, after one week of treatment, anesthetize with alcohol, kill, collect ovarian samples, freeze in liquid nitrogen, and store in -80 ℃ ultra-low temperature refrigerator save. The collected ovarian samples were extracted for total RNA, reverse-transcribed, and quantitatively detected by fluorescence to detect the expression changes of the ERR gene.

[0064] The result is as Figure 7 as shown in Figure 7 The results in showed that: compared with the control group, the mRNA exp...

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Abstract

The invention discloses a macrobrachium rosenbergii estrogen-related receptor gene ERR. The nucleotide sequence of the gene ERR is shown as SEQ ID NO:1, and the amino acid sequence of the gene ERR is shown as SEQ ID NO:2. The invention further discloses application of the recombined macrobrachium rosenbergii estrogen-related receptor gene ERR to environmental internal-secretion interfering-substance detection and pollution assessment, and macrobrachium rosenbergii ERR protein has important application value in the aspects of natural water environment internal-secretion interfering-substance (like NP) detection and pollution assessment.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and in particular relates to a macrobrachium rosenbergii estrogen-related receptor gene ERR and an application thereof. Background technique [0002] The estrogen-related receptors ERRα and ERRβ were first discovered by Giguere in 1988. For the conserved sequence of the DNA binding domain (DBD) of ERα, these two new human genes were screened by low-stringency hybridization method. They have a strong homology with the estrogen receptor (ER), hence the name. At present, three members of the ERR family have been found, namely ERRα (NR3B1), ERRβ (NR3B2) and ERRγ (NR3B3), which together with ER constitute a subfamily of the third class of nuclear receptors. Hormone binding, no response to the stimulation of natural estrogens such as estradiol, studies suggest that ERR can activate the expression of target genes by interacting with coactivators (coactivators) in the absence of ligands, so ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/12C12N15/70C07K14/72
CPCC07K14/43509C07K14/721
Inventor 朱春华陈华谱赵苗鑫李广丽邓思平洪越群洪宇键
Owner 海南永贺生物科技有限公司
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