Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

A method for regulating the period length of type I collagen fiber striations

A collagen fiber and collagen technology, which is applied in the field of genetic engineering, can solve the problems of high culture cost, long cycle, limited application and the like, and achieve the effect of simple preparation process and low cost

Active Publication Date: 2021-08-13
JIANGNAN UNIV +1
View PDF7 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The current sources of collagen are mainly the following five: the most important source is animal skin extraction, although the price is low, but it is easy to carry the source of disease; chemically synthesized peptides are highly controllable and high in purity, but the most obvious disadvantage is that they are expensive, Limited length, not suitable for mass production
The advantage of using eukaryotic systems such as transgenic plants and mammalian cells is that it can guide the correct folding of proteins and provide complex post-translational processing functions, but the common problems are high cost of culture, long cycle, low expression level and difficulty in scale Production
Recombinant collagen has potential applications in the production of biomaterials, but lacks the self-assembly driving force to form higher-order structures and cannot form higher-order structures, which limits its application in biomaterials and tissue engineering

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • A method for regulating the period length of type I collagen fiber striations
  • A method for regulating the period length of type I collagen fiber striations
  • A method for regulating the period length of type I collagen fiber striations

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] Example 1 Sequence design and sample preparation

[0052] according to The structure shown is designed, and the specific steps are:

[0053] (1) With N and C terminals (GPP) 10 As a fixed sequence motif, a variable collagen region is inserted in the middle to obtain a three-segment chimeric sequence (abbreviated as P 10 CLP 10 ). In this example, the CL-domain adopts the amino acid sequence (abbreviation is H) as bacterial collagen, where the Scl2 collagen domain is divided into three equal-length domains A, B and C. In the following examples, the designed CL domains are A, B, C, BB (two repeated B regions) and ABC (equivalent to the complete Scl2 collagen region).

[0054] (2) A globular domain derived from Scl2 (shown in SEQ ID NO.1) is inserted at the N-terminus of the sequence to guide the correct folding of the collagen triple helix, and a protease cleavage site is inserted between the globular domain and the fixed sequence unit of the collagen region LVP...

Embodiment 2

[0060] Embodiment 2 sequence design and sample preparation

[0061] The specific implementation method is the same as in Example 1, the difference is that the amino acid length of the CL domain is increased, and the designed CL domains are BB (repeated 2 B regions) and ABC (equivalent to the complete Scl2 collagen region), and the corresponding complete amino acids The sequence and nucleotide sequence are:

[0062] V-P 10 B 2 P 10 The amino acid sequence is shown in SEQ ID NO.10, encoding V-P 10 B 2 P 10 The gene sequence of is shown in SEQID NO.16.

[0063] V-P 10 ABCP 10 The amino acid sequence is shown in SEQ ID NO.11, encoding V-P 10 ABCP 10 The gene sequence of is shown in SEQID NO.17.

[0064] The proteins prepared in Examples 1-2 were identified. figure 2 (B) shows that a single band can be detected by SDS-PAGE for the digested protein. Since collagen is a rod-shaped protein, the protein marker used is a spherical molecule, and the molecular weight shown by...

Embodiment 3 2

[0065] Example 3 Secondary Structure Determination

[0066] The collagen prepared in Examples 1-2 was prepared at a concentration of 1 mg / mL. Then stand at 4°C for more than 24h, use a 1mm cuvette, scan the full wavelength of circular dichroism at 4°C, the wavelength is from 190nm to 260nm, the wavelength interval is 1nm, and stay at each wavelength for 5s. The thermal change experiment was measured at 220nm, the temperature was from 4°C to 80°C, each temperature was equilibrated for 8s, and the temperature increment rate was 1°C / 6min. The typical CD spectrum of collagen triple helix structure shows a positive absorption peak at 220nm.

[0067] Such as Figure 4 As shown, under full-wavelength scanning, the proteins designed in Examples 1 and 2 have characteristic absorption peaks around 220nm; the results of thermal variation experiments show that, with the increase of temperature, the characteristic absorption values ​​at 220nm are all abrupt at around 50°C. Changes, manife...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

PropertyMeasurementUnit
lengthaaaaaaaaaa
Login to View More

Abstract

The invention discloses a method for regulating the period length of type I collagen fiber striations, and belongs to the technical field of genetic engineering. The present invention uses N and C terminal (PPG) n Based on the sequence, a continuous collagen sequence with different numbers of Gly‑Xaa‑Yaa triplets is inserted in the middle to form a three-segment chimeric collagen P‑CL‑P pattern, through the N and C-terminal (PPG) n Interactions among the triple helices drive self-assembly to form ribbon-like fibers with periodic alternating light and dark stripes of varying dark stripe lengths. The method of the invention has a simple preparation process, can produce collagen fibers with low cost on a large scale, and has broad application prospects in the field of biomaterials.

Description

technical field [0001] The invention relates to a method for regulating the period length of type I collagen fiber stripes, and belongs to the technical field of genetic engineering. Background technique [0002] Collagen is a biological polymer, which is a triple helix structure formed by three chains intertwined with each other. Collagen can be divided into 28 types according to its gene sequence and function, the most important of which is type I collagen. In higher biological cells, after the original protein of type I collagen undergoes a series of maturation processes such as post-translational modification, folding, and cleavage, multiple collagen triple helices (collagen for short) are staggered to form light and dark stripes with uniform spacing. collagen fibers ( figure 1 ), whose morphology plays a key role in cell adhesion and growth [1] , is also a biomaterial that promotes the repair and regeneration of human tissues and organs. [0003] The current sources...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/78C12N15/12C12N15/70C12N1/21C12R1/19
CPCC07K14/78C12N15/70
Inventor 许菲胡金远维卡斯·南达戴维·史瑞伯张萌锁娜儿·格拉瓦特
Owner JIANGNAN UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com