Cloning and biological information analysis of alpha interferon from Yangzhou white goose

Abstract

Since ancient times, our country is a big country of poultry breeding, and our poultry breeding industry has a long history. At the same time, with the continuous development of China's livestock and poultry breeding industry in recent years, the national government has issued a series of relevant policies to support and regulate the development of the poultry industry, providing a good development environment for the poultry industry. However, with the development of large-scale intensive breeding industry, there are more and more diseases of geese, among which viral infectious diseases are the main cause of death of geese. Yangzhou white goose is the first local breed bred with domestic goose breed resources in China, and is known as the first new goose breed in China. Yangzhou White goose is an ideal goose breed because of its high reproductive rate, stable genetic performance, rapid initial growth of goslings for meat, and high muscle protein content. However, the breeding process of white geese in Yangzhou is often affected by a variety of infectious diseases, especially viral diseases such as bird flu, paramyxovirus disease and bursal disease of geese, which threaten the development of goose breeding industry. If a large number of chemical agents are used in the aquaculture industry, it will not only threaten people's health, but also lead to increased drug resistance of the virus, and interferon has the advantages of wider spectrum, high efficiency and safety than chemical agents. In this experiment, type I interferon alpha with strong antiviral activity was selected for the experiment, and the sequence-designed primer on goose interferon alpha published in Gen Bank was referred to. The interferon alpha gene was amplified and cloned by RT-PCR technology, and named yz goose IFNα-1 and yz goose IFNα-2. The nucleic acid and amino acid sequences of interferon alpha gene were analyzed by bioinformatics software and on-line analysis method. In this study, we obtained a fragment of 576bp size encoding 191 amino acids with a molecular weight of about 21.6kDa, which is consistent with the expected size. The obtained alpha-interferon sequences of two Yangzhou white geese were much more similar to those of chicken and duck than to those of human and other animals. The two sequences obtained above were imported into the online software NetPhos 2.0 for the prediction of phosphorylation sites. When the threshold was set at 0.5, the two amino acid sequences each contained 17 potential phosphorylation sites. The hydrophobicity of the known yz goose IFNα-1 and yz goose IFNα-2 derived peptide chains was analyzed by online ProtScale software. According to the hydrophobicity map, the maximum value of hydrophobicity of the two genes was 2.856, and the minimum value was -2.656. The first hydrophobic zone is located at 10-30aa and is the most hydrophobic.