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

Preparation method and use of fixed-length recombinant polynucleotide/polydeoxyribonucleotide

A deoxyribonucleotide and fixed-length technology, which is applied in the field of fixed-length recombinant polynucleotide/polydeoxyribonucleotide preparation, can solve the difficulty in controlling the size of PN/PDRN fragments, the risk of animal source contamination, and the difficulty in obtaining raw materials To achieve the effect of increasing conversion efficiency and unit yield, easy acquisition, and avoiding disease infection

Pending Publication Date: 2022-01-11
尚诚怡美(成都)生物科技有限公司
View PDF6 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0012] In view of the problems existing in the prior art, such as difficulty in obtaining raw materials, high cost, risk of animal source contamination, and difficulty in controlling the size of PN / PDRN fragments, the present invention proposes a method for preparing recombinant PN / PDRN with a fixed length and its application. To: provide a method that requires clear materials, short preparation time, and can prepare fixed-length PN / PDRN

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
  • Preparation method and use of fixed-length recombinant polynucleotide/polydeoxyribonucleotide
  • Preparation method and use of fixed-length recombinant polynucleotide/polydeoxyribonucleotide
  • Preparation method and use of fixed-length recombinant polynucleotide/polydeoxyribonucleotide

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0066] (1) Select a fixed-length PN / PDRN transformant from the PN / PDRN DNA library, called pUC18-PN / PDRN01, and the composition sequence of PN / PDRN01 is as follows:

[0067]

[0068]

[0069] (2) Culture pUC18-PN / PDRN01 transformants in Luria-Bertani medium containing 50 μg / mL ampicillin, and culture at 35±2°C for 16 hours;

[0070] (3) The pUC18-PN / PDRN01 recombinant cloning vector was extracted by alkaline lysis and alcohol precipitation;

[0071] (4) Digestion with BamHI+KpnI at 37°C for 4 hours, and the recombinant PN / PDRN01 can be excised from the recombinant cloning vector;

[0072] (5) Separating the pUC18 / BamHI+KpnI carrier cloning vector and the recombinant PN / PDRN01 by agarose gel electrophoresis, and using DNA purification resin to purify the recombinant PN / PDRN01 fragment and the pUC18 / BamHI+KpnI carrier cloning vector of a fixed length;

[0073] (6) Design and synthesize the KpnI-BamHI linker (linker), the sequence is as follows:

[0074] Forward 5'-CGGGGGG-...

Embodiment 2

[0082] (1) Select a fixed-length PN / PDRN transformant from the PN / PDRN DNA library, called pUC18-PN / PDRN02, and the composition sequence of PN / PDRN02 is as follows:

[0083]

[0084]

[0085]

[0086] (2) Culture pUC18-PN / PDRN02 transformants in Luria-Bertani medium containing 50 μg / mL ampicillin, and culture at 35±2°C for 16 hours;

[0087] (3) The pUC18-PN / PDRN02 recombinant cloning vector was extracted by alkaline lysis combined with alcohol precipitation;

[0088] (4) M13 sequencing primer sequence: 5'-GTAAAACGACGGCCAG-3' determines the recombinant PDRN02 sequence;

[0089] (5) Design and synthesize the primers required for polymerase chain reaction (primers). The principle of primer design is that the forward primer (forward primer) uses the sequence containing the BamHI restriction endonuclease recognition site on pUC18, and the reverse primer (reverse primer) is Use 10-12 nucleotides of the 3' end sequence on PDRN02 and add 5 sequences of BamHI restriction end...

Embodiment 3

[0100] (1) Select a fixed-length PN / PDRN transformant from the PN / PDRN DNA library, called pUC18-PN / PDRN03, and the composition sequence of PN / PDRN03 is as follows:

[0101]

[0102]

[0103] (2) The length of PN / PDRN03 should not be too long, preferably 50-60 base pairs;

[0104] (2) Culture pUC18-PN / PDRN03 transformants in Luria-Bertani medium containing 50 μg / mL ampicillin, and culture at 35±2°C for 16 hours;

[0105] (3) The pUC18-PN / PDRN03 recombinant cloning vector was extracted by alkaline lysis combined with alcohol precipitation;

[0106] (4) Sequence the primer sequence with M13: 5'-GTAAAACGACGGCCAG-3' determine the sequence of PDRN03;

[0107] (5) Design and synthesize the complementary sequence of the KpnI-PN / PDRN03-BamHI linker, the sequence is as follows:

[0108] forward sequence 5'- G PDRN03 sequence G -3'

[0109] reverse sequence 3'- GTACC PDRN03 sequence CCTAG -5' (underlined are the 5 sequences of KpnI and BamHI restriction endonuclease recog...

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

No PUM Login to View More

Abstract

The invention discloses a preparation method and use of a fixed-length recombinant polynucleotide / polydeoxyribonucleotide. The preparation method comprises the following steps: establishing a DNA library of PN / PDRN, selecting fixed-length PN / PDRN from the library, connecting to generate a multi-copy PN / PDRN fragment, and transferring the multi-copy PN / PDRN fragment to a cloning vector to obtain a recombinant PN / PDRN cloning vector; introducing the recombinant PN / PDRN vector into a suitable bacterial host to obtain a transformant, culturing the transformant to enable the recombinant PN / PDRN cloning vector to be massively copied and amplified, separating the recombinant PN / PDRN cloning vector from the transformant, and carrying out enzyme digestion from the recombinant PN / PDRN cloning vector to obtain a recombinant fixed-length PN / PDRN fragment. Different from a traditional method for extracting PN / PDRN from human or animal cell tissues, the method for preparing the recombinant fixed-length PN / PDRN has greatly improved production efficiency, avoids risks of human or animal cell tissue materials, has advantages of high yield and low cost, can be used in cosmetics, health-care products, medical devices, pharmaceuticals and the like, and obviously improves economic benefits.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for preparing a fixed-length recombinant polynucleotide / polydeoxyribonucleotide and an application thereof. Background technique [0002] As we all know, deoxyribose nucleic acid (Deoxyribonucleic acid, DNA) is the genetic material of most organisms including humans, responsible for the maintenance and transmission of life genetic information; DNA is a macromolecular polymer composed of four deoxyribonucleotides, deoxyribose Ribonucleotides are composed of deoxyribose sugar, bases and phosphates. There are four kinds of deoxyribonucleotides: adenine deoxyribonucleotide (Adenine deoxyribonucleotide, A), guanine deoxyribonucleotide (Guanine deoxyribonucleotide, G), thymine deoxyribonucleotide (Thyminedeoxyribonucleotide, T) and Cytosine deoxyribonucleotide (Cytosine deoxyribonucleotide, C). DNA molecular structure is composed of two reverse complementary polydeox...

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 Applications(China)
IPC IPC(8): C12N15/10C12N15/70
CPCC12N15/1093C12N15/10C12N15/70C12Q2565/125
Inventor 周江鸿李同琪徐震梅洪涛李朔
Owner 尚诚怡美(成都)生物科技有限公司
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