Mixed spidroin fiber preparing method

Abstract

The invention relates to a mixed spidroin fiber preparing method.According to the method, the araneus ventricousus minor ampullate spidroin full-length coding gene and a piriform silk complete repetition area are taken as a template, splicing is conducted with the fusion PCR technique, connection with a pLX carrier is conducted, recombination quality construction is conducted, and expression is conducted in BL21.The degeneration purification means is adopted, and a mixed spidroin solution is obtained finally and stored after freeze-drying.Reaction conditions are mild, cost is low, the process is simple, and operation is easy.Prepared mixed spidroin fiber has high biocompatibility, uniform thickness and a smooth surface, and has potential application value in fields like biomedicine.

Description

technical field [0001] The invention belongs to the field of preparation of protein fibers, in particular to a method for preparing mixed spidroin fibers. Background technique [0002] Spider silk is a natural high-molecular-weight protein fiber with good mechanical properties and biological qualities, such as high strength, good elasticity, light texture, high and low temperature resistance, and good biocompatibility. The overall quality far exceeds that of silk And the artificially synthesized high-performance material fiber at this stage is a strategic resource that needs to be developed urgently, and has inestimable application value in the fields of biomedical engineering, aerospace and military. However, the large-scale mass production and bionic applications of spider silk still need to be strengthened, mainly for the following reasons: 1. Spiders are carnivorous animals, which are not easy to domesticate and cannot be raised in high density like silkworms; 2. The com...

AI Technical Summary

Problems solved by technology

However, the large-scale mass production and bionic applications of spider silk still need to be strengthened, mainly for the following reasons: 1. Spiders are carnivorous animals, which are not easy to domesticate and cannot be raised in high density like silkworms; 2. The combination of natural spider silk 2. The silk forming mechanism and spinning process are extremely complex; 2. Using genetic engineering to express and produce recombinant spidroin in heterologous hosts is by far the most promising method

But this also faces difficulties, because the center of most spider silk proteins is a highly repetitive sequence, its molecular weight can reach more than 90%, and the repetitive region is rich in glycine and alanine

When expressing spider silk proteins with heterologous hosts, not only the problem of codon usage bias is faced, but this sequence characteristic also brings about the need for co-evolution of the library, and at the same time, this high content causes a special secondary structure and leads to gene recombination

Therefore, the produced spider silk protein reaches the size of natural spider silk protein, the yield is low, and the performance is far inferior to natural spider silk protein

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