Recombinant III-type human-like collagen as well as coding gene and expression method thereof

By optimizing the human type III collagen sequence and adapting to the codon characteristics of lettuce, expressing and extracting recombinant type III human collagen, the safety hazards and high cost problems in the existing technology and the extraction of animal-derived collagen are solved, and safe and efficient collagen production is achieved.

CN119684438APending Publication Date: 2025-03-25宋灵杰
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Patent Information

Application Number
CN202411943207.2
Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
Filing Date
2024-12-27
Publication Date
2025-03-25

AI Technical Summary

Technical Problem

In the prior art, when collagen is expressed by microorganisms, the translation expression efficiency is low due to the difference in codon preference and mRNA structure, and the extraction of animal-derived collagen from collagen is poor, safety hazards and high cost.

Method used

By optimizing the sequence of human type III collagen sequences, adapting to the codon properties of lettuce, eliminating enzyme cleavage sites and splicing sites that are unfavorable for expression, designing suitable PCR amplification primers, constructing recombinant plasmids and transforming Agrobacterium, and finally expressing and extracting recombinant type III human collagen in lettuce.

Benefits of technology

The expression efficiency and purity of recombinant type III human collagen is improved, the toxicity of protein is reduced, safe and efficient application in the field of biomedical science, and production costs are reduced.

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Abstract

The invention discloses a recombinant III-type human-like collagen as well as a coding gene and an expression method thereof, and the method comprises the following steps: S1, carrying out sequence optimization by taking a human III-type collagen sequence as a template to obtain a recombinant III-type human-like collagen gene sequence; designing PCR (Polymerase Chain Reaction) amplification upstream and downstream primers according to the gene sequence, and amplifying the gene sequence of the recombinant III type human-like collagen; s2, carrying out double enzyme digestion on the recombinant III type human-like collagen gene and a plasmid, and connecting to obtain a recombinant plasmid; s3, linearizing the recombinant plasmid, transforming the linearized recombinant plasmid into agrobacterium, and screening; and S4, infecting lettuce with agrobacterium, culturing to express protein, freezing the infected lettuce with liquid nitrogen, grinding, extracting total plant protein, and purifying to obtain the recombinant III-type human-like collagen. The recombinant III type human-like collagen is expressed through lettuce, so that the method is safe and efficient, and has sufficient market value and important practical significance.
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