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

Dopamine receptor new sub-type D5B gene and coded protein and use

A dopamine and gene technology, applied in the fields of genes and encoded proteins and applications, can solve the problem that the functions are not exactly the same

Inactive Publication Date: 2007-04-11
EAST CHINA NORMAL UNIV
View PDF0 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

D1 receptors and D5 receptors are located differently at the cellular and subcellular levels, suggesting that D1 and D5 receptors, while pharmacologically identical, are not functionally identical

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
  • Dopamine receptor new sub-type D5B gene and coded protein and use
  • Dopamine receptor new sub-type D5B gene and coded protein and use
  • Dopamine receptor new sub-type D5B gene and coded protein and use

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Cloning method of D5B1

[0032] Using the following primers, D5 was amplified by PCR using human genomic DNA as a template. The solid underline in the primers is the restriction site of EocRI, and the wavy underline is the restriction site of HindIII.

[0033] Primer 1: 5'-GGC GAA TTC GCG TGT GTG TGC GTG CTT GTC AGT GT-3’

[0034] Primer 2: 5'-GGG AAG CTT CTG AAG TTG GGA CCG CGC ACA GAC CG-3’

[0035] The reaction was carried out under the following conditions. Pre-denaturation at 94°C for 5 minutes; denaturation temperature at 94°C for 30s, annealing temperature from 50 to 60°C, one cycle every 2°C, and finally 20 cycles at 60°C, a total of 30 cycles, each cycle extended at 72°C for 2 minutes, and finally Extend at 72°C for 7 min.

[0036] The PCR product was purified by a PCR product purification kit (vitegene) to obtain a 0.025 μg / μl PCR product, and then co-digested with HindIII (TaKaRa) and EocRI (TaKaRa). The vector pcDNA3.1 was digested in the same way....

Embodiment 2

[0040] D5B cloning method 2

[0041] The genomic DNA of human SHSY5Y cells was used as a template, and the following two pairs of primers were used for PCR reaction.

[0042] Primer 1: 5′-gca gcc caa gcg gat cct gcg gat ctg cag tcc agc ccg aaa tgc t-3’.

[0043] Primer 2: 5′-gaa tag ggc cct cta gat gca tgc tcg agc gga tat ctt aat gga atc ​​cat tcc ggg t-3’

[0044] The PCR reaction was carried out under the following conditions, pre-denaturation at 94°C for 5min; denaturation temperature at 94°C for 30s, annealing at 55°C for 30s, extension at 72°C for 2min, 30 cycles. The PCR product was analyzed by 1% agarose gel electrophoresis, the PCR product of 1.5 kb was recovered, connected to the pMDT-18 vector (TaKaRa), and 41 clones were sequenced, of which 24 clones were D5 receptors, 12 One clone is D5B receptor, four clones are D5 pseudogene ψD51 and one pseudogene ψD52.

Embodiment 3

[0046] Gene transfection and cell culture

[0047] CHO cells were cultured at 37°C in RPMI1640 medium containing 10% fetal bovine serum, using cell transfection reagent (Lipofectin) reagent, with 6CRE response element, containing TATATA box (TA) minimized promoter and luciferase gene (Luciferase) constitutes the reporter gene, and the D5B gene and the reporter gene are transfected into CHO cells. The stable transgenic cells D5B / 6CRE / TA / Luci were screened in the medium containing 0.8mg / mL G418.

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 provides one new dopamine receptor sub-type D5B gene. The new dopamine receptor sub-type D5B gene obtained through PCR process is similar to dopamine receptor D5 and codes one G-protein coupling receptor containing 478 amino acids. Compared with dopamine receptor D5, the dopamine receptor D5B has even higher affinity to dopamine. The new sub-type D5B gene may be used as the important target point in developing medicine for psychosis, neural disease, cardiac vascular diseases and nephropathy.

Description

technical field [0001] The invention relates to the field of genetic engineering, in particular to a gene of a new dopamine receptor subtype, its encoded protein and its application. Background technique [0002] Some neuropsychiatric diseases (including: Parkinson's disease, schizophrenia, drug dependence, migraine, mania, depression and Tourette's syndrome, etc.), cardiovascular disease, kidney disease are related to dopamine receptors, and dopamine receptors are in the central nervous system Expressed in, control motor function, emotion, endocrine physiological system. [0003] Much research has been done on dopamine and where its receptors act. Cools and Rossum proposed through electrophysiological and pharmacological experiments that there may be more than one dopamine receptor in the brain, which is a milestone in dopamine research. In the 1970s, studies of dopamine receptors based on second-messenger assays, such as stimulation by cAMP, and binding assays supported ...

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): C12N15/12C07K14/435C12N15/63C12N5/10A61K48/00A61P25/00A61P9/00A61P13/12
Inventor 许治良江南胡应和
Owner EAST CHINA NORMAL 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