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A kind of bahd acyltransferase gene and its application

A technology of use and gene fragment, applied in the application field of BAHD acyltransferase gene, to achieve the effect of increasing value

Active Publication Date: 2021-05-28
AGRI RES INST TIBET ACADEMY OF AGRI & ANIMAL HUSBANDRY SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Highland barley also has certain medicinal value, but there is no report of feruloyltryptamine in highland barley in the prior art

Method used

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  • A kind of bahd acyltransferase gene and its application
  • A kind of bahd acyltransferase gene and its application
  • A kind of bahd acyltransferase gene and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1 Isolation and prokaryotic expression of HVUL2H29914 gene

[0034] This example is mainly about the method of HVUL2H29914 gene acquisition, vector construction and prokaryotic expression.

[0035] (1) Construction of gene fragments and vectors

[0036] Weigh 1 gram of fresh barley leaves, extract barley RNA, synthesize cDNA with M-MLV Reverse Transcriptase from Thermo Fisher Company, and design primers as follows:

[0037] F: CCGAGATCTATGAAGATCACCGTGCACTCTTC

[0038] R: CCGGAATTCCTAAAAGAGTGGCACGTAGGCGTCC

[0039] Perform PCR amplification to obtain fragments of the target band size (results such as figure 1 shown). The PCR product was purified with a gel extraction kit (Gel Extraction Kit D2500-02, OMEGA).

[0040] The nucleotide sequence (SEQ ID NO.1) of the amplified target fragment HVUL2H29914 gene is:

[0041]ATGAAGATCACCGTGCACTCTTCCAAGGCCGTCAAGCCCGAGTACGGCGCCTGCGGCCTCGCTCCGGGGTGTACCGCCGACGTCGTCCCGCTCACCGTGCTCGACAAGGCCAACTTCGACACGTACATCTCGGTGATCTACGCC...

Embodiment 2

[0057] The construction of embodiment 2 transgenic tobacco

[0058] ①Transform the transient expression vector containing the target gene (transient expression vector pEAQ, from John InnesCentre) into Agrobacterium (EHA105);

[0059] ②Pick positive Agrobacterium clones in 500ul LB containing corresponding antibiotics (kn), and culture them for 20-24 hours;

[0060] ③Transfer 200ul into 5ml LB containing the corresponding antibiotic (kn), shake at 28°C at 220rpm to about OD=2.0.

[0061] ④ Centrifuge at 10000rpm at room temperature for 2 minutes to collect the bacteria, resuspend the bacteria with the transformation buffer prepared in advance, and shake on the shaker for 3 hours; the composition and concentration of the buffer working solution are as follows: 10mM MES (pH5.7), 10mM MgCl2, 100μM acetosyringone (acetosyringone, AS).

[0062] ⑤Take the prepared 1ml syringe, remove the needle, select a syringe with a smooth outlet to inhale the bacterial liquid, take the Nicotian...

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Abstract

The invention provides a BAHD acyltransferase gene HVUL2H29914 derived from highland barley. The invention also provides a preparation method for a recombinant vector containing the gene, a recombinant bacterium, and a transgenic plant. The invention also provides a BAHD acyltransferase and a preparation method thereof . The BAHD acyltransferase of the invention can use feruloyl coenzyme A as an acyl donor and tryptophan as an acyl acceptor to synthesize feruloyl tryptamine, and has excellent application prospects.

Description

technical field [0001] The invention relates to the field of genetic engineering, in particular to the application of a BAHD acyltransferase gene derived from highland barley. Background technique [0002] Feruloyltryptamine has anti-inflammatory, anti-tumor and antibacterial effects, and is distributed in trace amounts in medicinal plants such as safflower. Due to limited medicinal resources, sources of feruloyltryptamine need to be further developed. [0003] Highland barley, English name of highland barley: hulless barley. It is a cereal crop of the genus Barley in the Poaceae family. Because of the separation of the inner and outer glumes and the bare kernels, it is also called naked barley, barley, and rice barley. It is mainly produced in China's Tibet, Qinghai, Sichuan, Yunnan and other places, and is the main food of the Tibetan people. [0004] Highland barley also has certain medicinal value, but there is no report in the prior art that highland barley contains ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/54C12N15/70C12N1/21C12N9/10A01H5/00A01H6/46A01H6/82C12R1/19
CPCC12N9/1029C12N15/8243
Inventor 巴桑玉珍王玉林原红军徐齐君曾兴权杨春葆于明寨扎桑韦泽秀顿珠加布尼玛扎西
Owner AGRI RES INST TIBET ACADEMY OF AGRI & ANIMAL HUSBANDRY SCI