Prokaryotic cell high-efficiency expression method of red rice heme binding protein

Abstract

The invention discloses a prokaryotic cell high-efficiency expression method of red rice heme binding protein, and relates to the technical field of prokaryotic cell expression, and the method comprises the following steps: preparing plant materials, carrying out sterile disinfection on red rice, carrying out incubator culture, taking a small amount of sterile red rice seedling samples, and extracting total RNA of the red rice by adopting a Trizol method; the method comprises the following steps: extracting total RNA (Ribonucleic Acid) of red rice, amplifying a target gene by RT-PCR (Reverse Transcription-Polymerase Chain Reaction), connecting the target gene with a recombinant expression vector, transforming escherichia coli cells by the recombinant vector, and inducing expression and detection of target protein escherichia coli. A target gene is connected with a recombinant expression vector, and then the recombinant vector is converted into escherichia coli cells, so that the high-efficiency expression of a target protein gene related to the binding of one heme in red rice in prokaryotic cell escherichia coli is realized; and key technical conditions and basis are provided for large-scale acquisition and activity identification and application of the plant-derived heme binding related protein.

Description

technical field [0001] The invention relates to the technical field of prokaryotic cell expression, in particular to a high-efficiency prokaryotic cell expression method of red rice hemopexin. Background technique [0002] Heme is one of the important members of the family of tetrapyrroles. With the deepening of research, it has been found that the functions of heme in human and mammalian cells are important and diverse. It is not only an important prosthetic group of hemoglobin, but also participates in the regulation of oxygen metabolism and transport, and intracellular electron transfer. In most cases, the non-free heme in the body often interacts with protein subunits to form a complex to exert its regulatory function. However, excessive free heme can induce the production of reactive oxygen species and cause cellular oxidative stress damage. Therefore, intracellular heme must be regulated by enzymes or specific proteins to maintain relative stability. Heme binding-r...

AI Technical Summary

Problems solved by technology

However, there are few studies on heme-binding related proteins in red rice, and their beneficial activities in vitro need to be further explored

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