Recombinant humanized III type collagen alpha 1 chain and application thereof

Abstract

The invention discloses a recombinant humanized III type collagen alpha 1 chain and an application thereof. The nucleotide sequence for coding the recombinant humanized III type collagen alpha 1 chainis shown as SEQID NO: 1. The recombinant humanized III type collagen alpha 1 chain sequentially comprises an amino-terminal affinity purification label, a humanized III type collagen mature peptide chain and a carboxy-terminal affinity purification label from an amino terminal. According to the recombinant humanized III type collagen alpha 1 chain disclosed by the embodiment of the invention, a specificity affinity purification label is designed at two ends of the humanized III type collagen mature peptide chain, so that purification of overall-length protein is facilitated, and the recombinant humanized III type collagen alpha 1 chain can also be used for identification of collagen. According to the recombinant humanized III type collagen alpha 1 chain disclosed by the embodiment of theinvention, the III type collagen mature peptide chain is adopted, the propeptide at two ends is removed, and the expression number of the gene without propeptide is far higher than those of the gene containing the propeptide.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, more specifically, the invention relates to a recombinant human type III collagen α1 chain and its application. Background technique [0002] Collagen is the main protein in all connective tissues including skin, bones, tendons and cartilage. For example, it forms large fiber bundles in the skin, each fiber bundle contains many individual collagen fibrils, making the skin structure strong and flexible. Collagen, a mature triple helix, is abundant in the human body and is valuable for a range of industrial and medical applications. Collagen has been widely used in clinical applications, and it has been proven safe and effective in soft tissue expansion, wound and burn repair, orthopedics and cardiovascular. [0003] Human type III collagen (hCOL3A1) is a fibril-forming collagen that is widely distributed in stretchable connective tissues such as skin, internal organs or vascular syst...

AI Technical Summary

Problems solved by technology

Moreover, the recombinantly expressed human collagen used for production usually does not add an affinity tag or only adds an affinity tag at one end, and the expressed mature peptide chain is easier to degrade. The protein purity obtained by traditional non-specific purification methods or single affinity tag purification Insufficient, it is difficult to remove some degradation products, and cannot meet the application with high purity requirements

In the prior art, the use of microorganisms to obtain collagen also has the following disadvantages: the DNA sequences expressing collagen are all natural coding sequences in the human genome (generally obtained by reverse transcription methods), but microorganisms and humans are involved in the process of transcription and translation of genes. There are certain differences among them, especially in the codon preference. Using the natural coding DNA sequence of human collagen to express in microorganisms will cause low translation and expression efficiency due to codon preference and mRNA structure.

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