Araneus ventricosus egg-case silk protein full-length gene and preparation method thereof

Abstract

The invention relates to an araneus ventricosus egg-case silk protein full-length gene and a preparation method thereof. The gene sequence is shown as SEQ ID NO.1. The preparation method comprises the following steps that a degenerate PCR (Polymerase Chain Reaction) method is used for amplifying NT and CT of the partial egg-case silk protein gene; the anchoring PCR is used for complementing the NT and the CT; a primer used for PCR amplification of the full length Tusp1 gene is designed according to the sequence obtained through sequence testing; a fragment obtained through amplification is subjected to TA cloning; the positive cloned bacteria comprising the full-length Tusp1 gene are subjected to plasmid extraction; full-length sequence testing is performed to obtain the araneus ventricosus egg-case silk protein full-length gene. The whole gene provided by the invention comprises the complete NT end, a repeated region and the complete CT end, and is used for coding the araneus ventricosus egg-case silk protein; the silk fiber has the better mechanical property and biocompatibility than silk and artificial fiber; in addition, no pollution is caused on the environment; the foundation is laid for the large-scale production of the araneus ventricosus protein in gene engineering.

Description

technical field [0001] The invention belongs to the field of spider silk proteins, and in particular relates to a full-length gene of spider silk protein and a preparation method thereof. Background technique [0002] Spider silk is a natural protein silk fiber with excellent mechanical properties, such as high strength, light texture, good elasticity, high and low temperature resistance, and good biological compatibility. Its various qualities are superior to silk and the current stage. Synthetic fibers. Due to its huge potential application value, it can be used in aerospace, military and biomedical fields in the future. But until now, the large-scale production and bionic applications of spider silk still face some challenges, mainly in: 1. Spiders are carnivores, live alone, and cannot be cultivated on a large scale like silkworms; 2. The components of natural spider silk and its silk formation The mechanism is still unclear; 3. The expression of spidroin in heterologo...

AI Technical Summary

Problems solved by technology

But until now, the large-scale production and bionic applications of spider silk still face some challenges, mainly in: 1. Spiders are carnivores, live alone, and cannot be cultivated on a large scale like silkworms; 2. The components of natural spider silk and its silk formation The mechanism is still unclear; 3. The expression of spidroin in heterologous hosts by genetic engineering is considered to be the most promising method so far

But there are also many difficulties. Because the size of the spidroin gene is usually more than 10kb, and there is a highly repetitive sequence in the center, only a few complete spidroin genes have been obtained.

Secondly, due to the fact that the repeat region is rich in glycine and alanine, as well as the problem of codon usage bias, the spidroin protein produced by gene recombination is not as good as the natural spidroin protein in terms of size, yield and performance.

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