A kind of preparation method and application of Sporothrix globosa hsp70_5 recombinant protein

Abstract

The invention discloses a method for preparing a recombinant protein of Sporothrix globosa HSP70_5 and its application. By extracting the total RNA of Sporothrix globosa, cDNA is obtained by reverse transcription PCR, and amplified by specific primers to obtain the heat-shock protein of Sporothrix globosa The full-length sequence of the protein 70 (HSP70) isoform 5 gene, using molecular biology methods to recombine the gene fragments in vitro and prepare the recombinant protein corresponding to the gene fragment, and apply the recombinant protein to rabbits to prepare the corresponding antibody. The antibody can be used to prepare an early rapid diagnosis product for Sporotrichum globosa infection. The present invention constructs the prokaryotic expression system of Sporothrix globosa HSP70_5 recombinant protein, which can be rapidly expressed in large quantities through induction; the purification process is simple and fast by using Ni affinity chromatography column; The antibody has good reaction specificity with the target protein.

Description

technical field [0001] The invention relates to the field of recombinant protein preparation. More specifically, the present invention relates to a preparation method and application of Sporothrix globosa HSP70_5 recombinant protein. Background technique [0002] Sporotrichosis (sporotrichosis) is the most common deep fungal disease caused by the biphasic dark fungus Sporothrix schenckii sensu lato (hereinafter referred to as Ss.). Ss. is a hyphae phase in the environment, parasitic or saprophytic in plants or soil; after infecting the organism, the lesion is a yeast phase. Ss. infection can form a chronic infection that is difficult to heal by itself, not only involving the skin, but also infecting organs through the lymphatic route, resulting in disseminated and systemic fatal infection. The three provinces in Northeast my country are one of the areas where the disease is most prevalent in the world. [0003] In the early days, people thought that S. schenckii was a sin...

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Problems solved by technology

Fungal culture also needs to use diseased tissue as a sample, and the culture result needs to wait for 7-14 days, which takes longer. At the same time, the growth of fungi is greatly affected by the environment. When the environmental temperature, humidity and other aspects are not suitable, it is not easy to grow. , so the positive rate cannot reach the ideal level we need

In addition, the above two methods are invasive examinations, and postoperative wound healing will leave scars, which are not ideal detection methods for children and patients with facial infections

As mentioned above, both fungal culture and pathological diagnosis take a long time, and medical institutions need to have high experimental conditions and professional technicians, which cannot be carried out in primary hospitals, and can only be treated with the clinical experience of doctors

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