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

New human-phosphoguanosine reductase, its coding sequence and application

A technology of guanosine phosphate and reductase, which is applied in the fields of biotechnology and medicine to achieve widespread effects

Inactive Publication Date: 2004-05-26
第二军医大学免疫学研究所
View PDF1 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The human GMPR gene was also cloned and expressed by Kanno et al., but the understanding of this gene has gone through a tortuous journey

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
  • New human-phosphoguanosine reductase, its coding sequence and application
  • New human-phosphoguanosine reductase, its coding sequence and application
  • New human-phosphoguanosine reductase, its coding sequence and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0092] Embodiment 1: Cloning of human GMPR2 cDNA

[0093] Total RNA of human dendritic cells was extracted with Trizol (Gibco). Then, poly(A) mRNA was isolated from total RNA. After the poly(A) mRNA was reverse-transcribed to form cDNA, the cDNA fragment was directional inserted into the multiple cloning site of the vector using the SuperScriptII cloning kit (purchased from Gibco), and transformed into DH5α bacteria to form a cDNA plasmid library. The 5' termini of randomly selected clones were sequenced by the dideoxy method. Comparing the determined cDNA sequence with the existing public DNA sequence database, it was found that the DNA sequence of one cDNA clone was a new full-length cDNA. The DNA sequence contained in the new clone is bidirectionally determined by synthesizing a series of primers. Computer analysis showed that the full-length cDNA contained in the clone was a new cDNA sequence (shown in SEQ ID NO: 1 and Figure 1), encoding a new protein (shown in SEQ ID ...

Embodiment 2

[0096] Embodiment 2: clone the coding sequence of human GMPR2 with RT-PCR method

[0097] Use Trizol (Gibco company) to extract the total RNA of B lymphoma cell line Raji cells in the logarithmic growth phase, take 6 mg of total cell RNA and 0.5 μg Oligo-dT 12-18 Mix and perform reverse transcription. The reverse transcription system is 20μl, add 80μl ddH after the reaction 2 O for dilution. The primers used in PCR amplification are as follows: sense primer 5'-G GAATTC CGCCATCATGCCTCATATTGACAACGATG-3' (SEQ ID NO: 3), antisense primer 5'-G GAATTC GTCGACGCACGCCTCACTGAAGATTG-3' (SEQ ID NO: 4). The PCR reaction volume was 50 μl, which contained 10 μl of reverse transcription template, 0.4 mM primers, 0.2 mM dNTP and 1U ExTaq DNA polymerase (Takara Inc.), and the amplification parameters were 94°C for 30 seconds, 60°C for 30 seconds, and 72°C for 45 seconds. Seconds, after 25 cycles, the PCR product was initially confirmed by 2% agarose gel electrophoresis. The result of DN...

Embodiment 3

[0098] Example 3 Northern blot analysis of GMPR2

[0099] Perform Northern blotting according to the following conventional method: put the filter membrane to be tested in 10ml of hybridization solution preheated at 68°C, and pre-hybridize at 68°C in a hybridization oven (Bellco) for 30 minutes; put the labeled cDNA probe at 95-100 Denature at ℃ for 2-5 minutes, put it on ice to cool quickly, and then add hybridization solution (the final concentration of cDNA probe is 2-10ng / ml or 1-2×10 6 cpm / ml), mix thoroughly, and hybridize at 68°C for 2 hours. After the hybridization, the filter membrane was washed several times with 2×SSC and 0.05% SDS at room temperature, followed by shaking and washing for 30-40 minutes, during which the washing solution was changed several times. Then rinse with 0.1×SSC and 0.1% SDS at 50° C. for 20 to 40 minutes with shaking. Finally, the filter membrane was wrapped with plastic cling film, and exposed to X-ray film at -70°C for 24 to 48 hours.

...

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 present invention relates to a novel human guanosine monophosphate reductase GMPR2. Said invention provides the polynucleotide for coding said protein molecule and method for producing this protein molecule by means of DNA recombination technology. It also discloses the application of the polynuleotide coding this novel guanosine monophosphate reductase, dand the zymological acitivity and the relationship of it and tumour cell multiplication and cell differentiation are verified. Said invention also discloses the strategy of resisting said protein molecule for diagnosing and curing diseases, speciallyf ro diagnosing and curing the diseases of tumour, etc.

Description

technical field [0001] The present invention belongs to the fields of biotechnology and medicine, in particular, the present invention relates to a new polynucleotide encoding human guanosine monophosphate reductase (guanosine monophosphate reductase 2, referred to as "GMPR2") polypeptide, and the polynucleotide encoding of polypeptides. The present invention also relates to the use and preparation of such polynucleotides and polypeptides. The invention also relates to the enzymatic activity of GMPR2 and its relationship with tumor cell proliferation and differentiation. Specifically, the polypeptide of the present invention is a novel intracellular protein related to cell proliferation, differentiation, apoptosis, and inflammatory response. Background technique [0002] Enzymes are a class of biomacromolecular substances with catalytic activity and special spatial conformation in human cells, which play an important role in maintaining normal intracellular substance metab...

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): A61K38/44A61P31/00A61P35/00C07H21/00C07K16/18C12N9/02C12N15/53C12N15/63C12N15/64C12Q1/26
Inventor 张嘉章卫平万涛曹雪涛
Owner 第二军医大学免疫学研究所
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