Promoter sequence of garlic allinase gene

A promoter sequence and alliinase technology, applied in the field of endogenous enzyme (alliinase) promoter sequence, can solve complex problems and achieve high specificity and high sensitivity

Pending Publication Date: 2020-06-16
XUZHOU NORMAL UNIVERSITY
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Problems solved by technology

For gene families, the study of genetic diversity, or h...

Method used

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Embodiment Construction

[0013] 1. Extraction of Garlic RNA

[0014] RNA was extracted and purified using RNA extraction kit. Reverse transcription was performed by a reverse transcription kit, and multiple primers were designed using Primer5.0 according to the alliinase CDS sequence (Accession: KR270349.1) published in the NCBI database. Primers synthesized by Shanghai Bioengineering Company were diluted for later use and stored in a -80°C refrigerator.

[0015] two. Using the primers shown in SEQ ID NO.2, 3, and 4 and the Genome Walking kit kit, various conditions such as cycles and temperatures that are conducive to the amplification of gene fragments were designed to carry out PCR reactions, and multiple experiments were carried out in the middle (AP1, AP2 , AP3, AP4 have been tested), and finally a mature PCR system was obtained, as follows:

[0016] 1st reaction:

[0017] Select an appropriate amount of cDNA as a template, use AP4 as an upstream primer, and SP1 as a downstream primer. SP1 is...

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Abstract

The invention relates to a garlic alliinase gene promoter sequence which consists of 2407 nucleotides in SEQ ID NO.1. The gene sequence of the alliinase promoter disclosed by the invention can providea powerful molecular basis for researching alliinase interaction genes and researching an allicin synthesis mechanism in the future. Defects caused by traditional PCR in alliinase promoter gene cloning are overcome, and by means of chromosome stepping, the the advantages of being efficient, easy and convenient to implement, high in specificity and sensitivity and long in unknown sequence are obtained at a time. The gene sequence of the alliinase promoter can provide a powerful molecular basis for researching alliinase interaction genes, researching an allicin synthesis mechanism and increasing the allicin content in the future.

Description

technical field [0001] The present invention relates to gene cloning in the field of gene molecular technology, specifically, the promoter sequence of an endogenous enzyme (alliinase) that affects the synthesis of allicin in garlic has been cloned. The present invention is for better research in the future. The alliinase gene interaction, the mechanism of allicin synthesis and the increase of allicin content provide a strong molecular basis. Background technique [0002] garlic( Allium sativum L.), Liliaceae Allium, is a medicinal and edible plant with a long history. As early as more than 4,000 years ago, people began to eat garlic and use garlic to treat various diseases. It has high medicinal value, and because of its unique The flavor traits are deeply loved by people. In my country, garlic has been cultivated for 2,000 years, and the production areas are mainly concentrated in Shandong, Jiangsu, Henan, Hubei, Shaanxi and other provinces. Now my country has become the...

Claims

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Application Information

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IPC IPC(8): C12N15/113
CPCC12N9/88C12N2830/34C12Y404/01004
Inventor 杨绪勤苏奕人王军娟吴家莹范飞陈让让徐玲霞沈紫璇
Owner XUZHOU NORMAL UNIVERSITY
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