A kind of Siniperca tlr7 gene and its application

A technology of mandarin mandarin fish and genes, which is applied in aquaculture and biological fields, can solve problems such as the report on gene expression research of TLR7 cloning pathway of mandarin mandarin fish, and achieve the effect of improving economic benefits

Active Publication Date: 2020-04-10
SUZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the cloning and expression of TLR7 genes in fishes such as Atlantic salmon, large yellow croaker, and flounder have been reported, but there are no reports on the cloning of TLR7 and the study of gene expression in related pathways.

Method used

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  • A kind of Siniperca tlr7 gene and its application
  • A kind of Siniperca tlr7 gene and its application
  • A kind of Siniperca tlr7 gene and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] 1. Cloning of TLR7 cDNA of Siniperca sinensis

[0026] The primers used for TLR7 gene cDNA amplification are shown in Table 1 below:

[0027] Table 1 The sequences of primers used in TLR7 gene cDNA amplification

[0028] Primer name Primer sequence (5'-3') Purpose GSP-TLR7-1 CAGTCCACCTCTCACCA TLR7 5'-Race GSP-TLR7-2 CTGCCGTTGCTGGTCACA TLR7 5'-Race GSP-TLR7-3 ATGGCAGGGTTTTTGGGT TLR7 5'-Race 3' CDS Primer A AAGCAGTGGTATCAACGCAGACTAC TLR7 3'-Race GSP-TLR7-4 GAAAGTGCCTTGCAATTCAAAGTACCTA TLR7 3'-Race GSP-TLR7-5 GTATAAGCGGTCTGTGCTGGAGTGG TLR7 3'-Race AUAP GGCCACGCGTCGACTAGTAC Universal 5'-Race AAP GGCCACGCGTCGACTAGTAC(G) 16

Universal 5'-Race UPM CTAATACGACTCACTATAGGGC Universal 3’-Race

[0029] 1. Amplification of Siniperca sinensis TLR7 5' end sequence

[0030] (1) Synthesis, purification and tailing of the first strand of cDNA

[0031] Use SUPERSCRIPT II RT enzyme and pr...

Embodiment 2

[0087] Infectious spleen and kidney necrosis virus (ISKNV) and Aeromonas hydrophila (Aeromonas hydrophila) are the most serious virus and bacterial pathogens in mandarin fish culture. Renal necrosis virus and Aeromonas hydrophila) were tested for real-time detection of TLR7-Myd88-dependent disease resistance signaling pathway-related genes (TLR7, Myd88, IRAK1 and The change of IRF7) expression is an auxiliary detection method for Siniperca sinensis infected by virus or bacteria.

[0088] Experimental method: A total of 100 healthy mandarin ducks with an average weight of 42.63g were placed in an aquarium with a water flow system for two weeks before the experiment began. The breeding conditions were continuous aeration, water temperature 26°C, Feed three times. Before the infection experiment, 10 fish were randomly selected to check the common pathogens such as infectious spleen and kidney necrosis virus (ISKNV) and Aeromonas hydrophila in the body, and after ensuring that th...

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Abstract

The invention discloses a Siniperca chuatsi TLR7 gene and an application thereof. As shown in SEQ ID NO: 1, that nucleotide sequence of the TLR7 gene of Siniperca chuatsi of the invention is use for detecting the initial pathogen infection of Siniperca chuatsi, in particular, the nucleotide sequence of the TLR7 gene of the Siniperca chuatsi of the invention is as follow: Myd88-dependent disease resistance signaling pathway-related gene design specific amplification primers, wherein, the TLR7-Myd88-dependent signaling pathway related genes include TLR7, Myd88, IRAK1 and IRF7 genes. QPCR is usedto detect TLR7 in Siniperca chuatsi at the early stage of infection. The significant response of Myd88-dependent signaling pathway related gene mRNA expression is detected and analyzed, and the detection results are used as an auxiliary index in the early stage of fish infection by virus or bacteria. The invention provides a new idea for early diagnosis of virus or bacterial infection of Siniperca chuatsi, which is favorable for early prevention and control of fish diseases and improves the economic benefit of mandarin fish culture.

Description

technical field [0001] The invention belongs to the fields of aquaculture and biotechnology, and in particular relates to a TLR7 gene of mandarin fish and its application in detection of mandarin mandarin fish at the initial stage of pathogen infection. Background technique [0002] Siniperca chuatsi, belonging to Perciformes, Sinipercinae, and Siniperca, is a valuable economic fish and an important freshwater aquaculture species in my country. However, with the continuous expansion of the scale of artificial breeding in recent years, the germplasm degradation and disease resistance of the mandarin fish are relatively common, leading to the increasingly rampant exposure of viral diseases, and there is no very effective control method so far. All cause serious economic losses every year, seriously restricting the sustainable development of its breeding industry. Therefore, the early detection and diagnosis of pathogenic infection of Siniperca sinensis is an important means t...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/29C07K14/415C12Q1/6888
CPCC07K14/415C12Q1/6888C12Q2600/158
Inventor 黄鹤忠路瑶叶金明肖攀李泽金锐铭董同瑚
Owner SUZHOU UNIV
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