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

Tropane alkaloid transport protein AbTAUP1 and application thereof

A technology for transporting proteins and alkaloids, applied in applications, biochemical equipment and methods, plant peptides, etc., can solve the problems such as no reports on tropane alkaloid transporters, and achieve the effect of increasing content and increasing yield

Active Publication Date: 2021-10-29
SOUTHWEST UNIVERSITY
View PDF2 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But unfortunately, there is still no relevant report on the tropane alkaloid transporter

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
  • Tropane alkaloid transport protein AbTAUP1 and application thereof
  • Tropane alkaloid transport protein AbTAUP1 and application thereof
  • Tropane alkaloid transport protein AbTAUP1 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1. Tissue expression pattern analysis and screening of potential tropine alkaloid transporters

[0025] (1) Transcriptome analysis to screen potential tropine alkaloid transporters

[0026] The Purine nucleobase transmembrane transporter (PUNUT) family widely exists in plants and fungi, and is involved in the transport of plant metabolites such as cytokinins, caffeine and nicotine. The corresponding Hidden Markov Model (HMM) accession number in the Pfam database is PF16913.5. Using the PF16913.5 model, 19 candidate genes were identified in the Belladonna transcriptome. Alkaline synthesis pathway enzyme genes were analyzed by tissue expression clustering, and a highly expressed PUNUT family transporter gene in roots was obtained, which was highly correlated with the expression of tropane alkaloid synthesis pathway enzyme genes. Base transport, named as belladonna-tropane alkaloid transporter 1 (Tropane Alkaloid Uptake Permease 1, AbTAUP1).

[0027] (2) qPCR to ...

Embodiment 2

[0039] Example 2, analysis of subcellular localization of AbTAUP1

[0040] (1) Construction of subcellular localization vector

[0041] The prediction of the transmembrane domain of the AbTAUP1 protein found that the protein contains ten transmembrane domains, and almost the entire protein runs through the membrane structure, so the AbTAUP1 protein is likely to exert the ability to transport substrates on the cell membrane. In order to verify that AbTAUP1 protein is a transmembrane protein, a subcellular localization vector was constructed to analyze the subcellular localization of AbTAUP1. The purchased pm-ck / CD3-1001 plasmid (purchase link https: / / abrc.osu.edu / stocks / 764634) can express the plasma membrane localization protein PM-CFP fused with cyan fluorescent protein CFP as a plasma membrane marker signal . After transforming Agrobacterium GV3101, the engineering strain GV3101-pCD3-1001 was obtained.

[0042] By analyzing the restriction enzyme sites in the coding regio...

Embodiment 3

[0048] Example 3, the application of AbTAUP1 in yeast

[0049] (1) Construction of yeast transfer vectors and engineering strains

[0050] To study whether AbTAUP1 gene has the ability to transport tropane alkaloids by constructing yeast transport vector. According to the multiple cloning site of the yeast expression vector pDR196, the upstream and downstream primers with PstI and XhoI restriction endonuclease cutting sites were designed. The specific primers are as follows:

[0051] pDR-AbTAUP1-F: 5'-ggg ctgcagatggaatctcaaattgctag-3' (SEQ ID NO.11);

[0052] pDR-AbTAUP1-F: 5'-ccc ctcgagttatattggaggggattgag-3' (SEQ ID NO. 12).

[0053] After PCR amplification, the pDR196 plasmid and the PCR product were double digested and recovered, then ligated, and finally the recombinant plasmid pDR196-AbTAUP1 was successfully obtained.

[0054] Yeast cells were transformed by chemical method, and the pDR196-AbTAUP1 recombinant plasmid was transferred into the Saccharomyces cerevisiae ...

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
wavelengthaaaaaaaaaa
Login to View More

Abstract

The invention discloses a tropane alkaloid transport protein AbTAUP1 and application thereof. An amino acid sequence of the tropane alkaloid transport protein AbTAUP1 is as shown in SEQ ID NO. 8, or an amino acid sequence which is obtained in a manner that the sequence as shown in the SEQ ID NO.8 is subjected to substitution, mutation or deletion of one or more amino acid residues and is encoded with the same function. The expression level of the protein in lateral roots is highest, the expression level of the protein in main roots is relatively low, the protein is located in a cytoplasmic membrane and speculated to have a hyoscyamine transport function, the hyoscyamine transport function of the protein is verified through a yeast transport experiment, the yield of hyoscyamine in atropa belladonna is increased by overexpressing the AbTAUP1 in atropa belladonna plants, so that the AbTAUP1 can be used for providing hyoscyamine transport capacity in hyoscyamine synthesis plants and increasing the hyoscyamine yield and has an important significance in increasing the content of tropane alkaloids in plant species.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the tropane alkaloid transporter AbTAUP1, and also relates to the application of the tropane alkaloid transporter. Background technique [0002] Tropane alkaloid (Tropane Alkaloid, TA) is a class of anticholinergic drugs acting on the parasympathetic nerve. Its representative drugs hyoscyamine, scopolamine and their derivatives have been widely used in anesthetics, treatment of Parkinson's, various organ colic and asthma caused by smooth muscle spasm, and the market demand is huge. At present, tropane alkaloids still completely rely on the extraction from a few medicinal plants of Solanaceae, including belladonna (Atropa belladonna), datura (Datura stramonium) and hyoscyamus niger. Belladonna is a commercially cultivated drug source plant for the production of hyoscyamine and scopolamine included and stipulated in the Chinese Pharmacopoeia. The mass fraction of scopolamine in wild ...

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): C07K14/415C12N15/29C12N15/82C12N15/81C12N1/19A01H5/00A01H6/82
CPCC07K14/415C12N15/8243C12N15/81
Inventor 廖志华陈敏杨春贤曾俊岚邱飞
Owner SOUTHWEST UNIVERSITY
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