Check patentability & draft patents in minutes with Patsnap Eureka AI!

Refractory phosphotriose isomerase gene and its polypeptide coded by it and preparing process

An isomeric, triose phosphate technology, applied in the field of genetic engineering, can solve the problems such as the lack of effective treatment of triose phosphate isomerase deficiency and the lack of effective research on the possibility of replacement therapy.

Inactive Publication Date: 2002-08-28
HUADA GENE RES & DEV CENT HANGZHOU
View PDF0 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the first cases occurred more than 30 years ago, until now there is no effective treatment for triose phosphate isomerase deficiency
The possibility of replacement therapy for this enzyme has not been effectively studied

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
  • Refractory phosphotriose isomerase gene and its polypeptide coded by it and preparing process
  • Refractory phosphotriose isomerase gene and its polypeptide coded by it and preparing process
  • Refractory phosphotriose isomerase gene and its polypeptide coded by it and preparing process

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1: Construction of a sequencing library

[0032] The sequencing library was constructed using the genome-wide shotgun method. Firstly, Tengchong thermophilic anaerobic bacteria were cultured according to (Yanfen Xue, 2000) modified MB medium (Balch et al., 1979), bacteria were collected according to Marmur (1961) method, and total DNA was extracted. In order to ensure the randomness of sequencing library construction and avoid the problem of hot spots of breakage to the greatest extent, a variety of methods and principles of library construction under different conditions were adopted. Firstly, physical shearing method (including ultrasonic method and shearing with Hydroshear Machine) was used, and then AluI was selected for random partial enzyme digestion according to the genome characteristics of the bacteria. Different intensities were used to process samples during physical shearing, and samples were processed by setting enzyme gradients during enzyme diges...

Embodiment 2

[0033] Example 2: Genome Sequencing

[0034] When sequencing the genome of thermophilic anaerobic bacteria in Tengchong, two automatic sequencers were mainly used: ABI377 and MegaBACE 1000. These two sequencers use the principle of electrophoresis for sequencing (see Figure 2), and can complete 96 samples each time. ABI377 is a product of ABI company, which is a kind of ABI series. It belongs to the slab gel electrophoresis sequencer. MegaBACE 1000 is a product of Pharmacia, which belongs to capillary gel electrophoresis sequencer.

Embodiment 3

[0035] Example 3: Data Analysis

[0036] 1) Basecalling and sequencing quality monitoring

[0037] The so-called basecalling refers to the process of obtaining the correct base sequence from the raw data file obtained on the sequencer. Since the sequencer obtains the intensity change traces (traces) of light of different wavelengths corresponding to the four bases A, T, G, and C, it is necessary to use a computer to adopt a certain algorithm to correctly identify the bases corresponding to the different traces. . We used Phred software (Ewing B, Hillier L, 1998) because its results are more reliable and its output is more convenient for further analysis by other programs in the same software package.

[0038] The algorithm principle of Phred's basecalling is to judge the base type based on the shape, distance, and signal-to-noise ratio of each peak in the trajectory, and at the same time give the credibility information for this base, that is, the sequencing quality of the b...

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

A refractory phosphotriose isomerase gene, the polypeptide coded by it and its preparing process are dislcosed. On the basis of full-genom sequencing and analyzing of a thermophilic anaerobe, the said refractory phosphotriose isomerase gene is cloned and separated. It can be used to prepare the transgenic microbes or animal or plant for producing the said gene and recovering the enzyme coded by the said gene. The amino acid sequence of the polypeptide with activity of the said gene, the function-equal object and a process for prepairng, separating and purifying the said polypeptide are also disclosed.

Description

technical field [0001] The invention relates to the field of genetic engineering, in particular to a high-temperature-resistant triose phosphate isomerase gene and its encoded polypeptide and a method for preparing the polypeptide. Background technique [0002] The triosephosphate isomerase (triosephosphate isomerase) involved in the present invention is one of the enzymes in the glycolysis pathway, and its function is to catalyze the conversion of dihydroxyacetone phosphate into 3-glyceraldehyde phosphate. The molecular weight of triose phosphate isomerase is 27261 Daltons, which is composed of 8 parallel β strands and 8 α helical strands surrounding the central core. Phosphoric acid is a weak competitor in this reaction, but when phosphoric acid and glycidol form phosphoglycidol, the reaction is strongly inhibited. [0003] Triose phosphate isomerase is an enzyme that widely exists in nature, and its role is to catalyze the exchange reaction of dih...

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
IPC IPC(8): C12N9/90C12N15/61C12N15/63C12N15/64
Inventor 包其郁汪建张小英胡松年李志杰
Owner HUADA GENE RES & DEV CENT HANGZHOU
Features
  • R&D
  • Intellectual Property
  • Life Sciences
  • Materials
  • Tech Scout
Why Patsnap Eureka
  • Unparalleled Data Quality
  • Higher Quality Content
  • 60% Fewer Hallucinations
Social media
Patsnap Eureka Blog
Learn More

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

© 2025 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com