Mosquito fish nrf2 gene and its extraction method and specific primary antibody preparation method
A mosquito-eating fish, specific technology, applied in DNA preparation, genetic engineering, plant genetic improvement, etc., can solve the problems of unclear genetic background of mosquito-eating fish, not including the whole genome, etc.
Active Publication Date: 2022-03-04
JINAN UNIVERSITY
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Problems solved by technology
[0005] The technical problem to be solved by the present invention is that the genetic background of mosquito fish is not clear, and the sequence of its whole genome is not included in the NCBI database
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[0050] The invention provides a mosquito fish Nrf2 gene extraction method and a specific primary antibody preparation method, which are convenient to operate and obtain the mosquito fish Nrf2 gene and specific primary antibody. The present invention will be described in detail below in conjunction with the accompanying drawings and specific embodiments.
[0051] The mosquito fish Nrf2 gene provided by the present invention is characterized in that the sequence of the gene fragment is as shown in Nrf2 ID NO.1.
[0052] A kind of mosquito fish Nrf2 gene extraction method provided by the invention is characterized in that, comprises the following steps:
[0053] 1) Extracting total RNA from the liver of mosquito fish;
[0054] 2) Obtain the first-strand cDNA of mosquito fish by reverse transcription according to the total RNA of mosquito fish liver;
[0055] 3) Using the first-strand cDNA of mosquito fish as a template, carry out ordinary PCR amplification to obtain the core fr...
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Abstract
The invention discloses a mosquito-eating fish Nrf2 gene, an extraction method thereof, and a method for preparing a specific primary antibody. The core base sequence of the Nrf2 gene in the mosquito-eating fish was successfully cloned for the first time, and the Nrf2-specific Nrf2-specific antibody was synthesized for the first time. Polyclonal primary antibodies can be widely used to quickly obtain unknown base sequences of other non-model species and prepare related specific antibodies, and use these genes to construct molecular-level detection methods and use antibodies to detect related protein changes in subsequent research. The mosquito-eating fish is highly adaptable to the environment, and can even live normally in sewage and sewers with high biological oxygen demand and chemical oxygen demand. It is speculated that its super vitality is also related to the Nrf2 antioxidant signaling pathway in its body. Methods The cloned gene and the prepared primary antibody can provide new technical means and detection methods for studying how Nrf2-related pathways play an antioxidant role under environmental stress and how organisms resist oxidative stress and related environmental risks.
Description
technical field [0001] The invention relates to the technical field of gene extraction and specific antibody preparation, in particular to a mosquito fish Nrf2 gene and its extraction method and a specific primary antibody preparation method. Background technique [0002] The mosquitofish (Gambusia affinis) is a small freshwater fish native to South America and belongs to the genus Gambusia affinis. The body size of female and male fish is obviously different: the body size of female fish is generally larger than that of male fish, the anal fin of female fish is fan-shaped, and the anal fin of male fish is sword-shaped. Mosquito fish is also known as potbelly fish and wicker fish, and its reproduction method is ovoviviparous. Mosquitofish have been widely introduced around the world to control mosquito-related outbreaks due to their strong environmental adaptability and broad-spectrum feeding habits. In recent years, mosquitofish, as an invasive species, has widely existed...
Claims
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IPC IPC(8): C12N15/12C12N15/10C07K16/18C07K16/06
CPCC07K14/461C07K16/18C07K16/065
Inventor 鲍爽聂湘平
Owner JINAN UNIVERSITY