DNA barcode, primers, kit and method, and applications thereof

A barcode and primer pair technology, applied in the field of kits, DNA barcodes and primers, can solve the problems of lack of Pu'er tea, damage to the economic interests of Pu'er tea production enterprises, etc. Effect

Active Publication Date: 2019-02-26
MENGHAI TEA IND +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In recent years, due to the increasing market demand, many small-scale manufacturers lack systematic and in-depth research on Pu-erh tea, and wantonly abuse many other people's patents for profit. A series of methods, such as Pu’er tea processing and production, have greatly damaged the economic interests of some Pu’er tea production enterprises with relevant patents.

Method used

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  • DNA barcode, primers, kit and method, and applications thereof
  • DNA barcode, primers, kit and method, and applications thereof
  • DNA barcode, primers, kit and method, and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0072] Example 1: Obtaining of LipB gene and DNA barcode

[0073] The proteome and genome analysis methods and mass spectrometry methods used in this example are all known methods in the art, so the specific operation process will not be repeated.

[0074] 1. Using the high-coverage proteome technology, pFind and pAnno software were used to conduct a deep coverage study of the proteome of the industrial fermentation fungus Arthrospora adenorivorum TMCC 70007, and its genome was verified for its annotated coding genes. Specifically, in order to discover new protein coding regions, using the Six Frame Translation strategy in systematic proteomics, a six-frame translation database of the genome data of Arthrospora adenophagous TMCC 70007 was obtained, exhaustively enumerating There are 6 coding possibilities of the genome (+1, +2, +3, -1, -2, -3), and the protein sequence is called "six-box translation protein sequence", and the corresponding nucleic acid sequence is called "six-...

Embodiment 2

[0093] Example 2. Identification of bacterial strains using DNA barcodes

[0094] According to the amplification result of the sample to be tested and the sequence homology with Arthrospora adenophagous TMCC 70007 strain SEQ ID NO.1, it is determined whether the sample to be tested is the industrially applied strain of Arthrospora adenorivorum TMCC 70007 .

[0095] (1) Based on SEQ ID NO.1, use the NCBI primer design tool to design PCR primers at both ends of the gene. The amplified product must include the start and end sites of the gene, and the sequences of the forward and reverse primers are respectively LipB-F: 5 '-AACCGGTCTCCGTCAAACTC-3'; LipB-R: 5'-AAGATCTCCGCGACAAGCAA-3'. For primer locations see Image 6 , the theoretical amplification length of the pair of primers is 1579bp, and the amplified sequence is SEQ ID NO.7.

[0096] (2) Source of strain

[0097] Table 4 Selected relevant strain information

[0098]

[0099] (3) Separately extract strain DNA: OMEGA E...

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Abstract

The invention specifically relates to a DNA barcode, primers, a kit and a method for identifying a strain of Blastobotrys adeninivorans, and applications thereof. The DNA barcode can accurately identify a Pu'er tea fermentation strain, i.e., Blastobotrys adeninivorans TMCC 70007, and can quickly and accurately identify the strain from confusing strains or other strains of the same species.

Description

technical field [0001] The invention belongs to the field of species and bacterial strain identification, and specifically relates to DNA barcodes, primers, kits, methods and applications. Background technique [0002] Pu-erh tea is a post-fermented tea with the geographical indication of Yunnan. It uses large-leaf sun-dried green tea as raw material and is made through a series of processes. The traditional production process of Pu'er tea is as follows: the picked fresh tea leaves are rolled, sun-dried, impurity-removed, tide-watered, heaped, dried, sieved, pressed and molded, and packaged for delivery. In the production of Pu-erh tea, the heaping fermentation process is the main factor to form the quality of Pu-erh tea. During this process, the internal components of the tea such as tea polyphenols, caffeine and some polysaccharides have undergone tremendous changes, making Pu-erh tea The special flavor, taste, quality and various health benefits of tea. [0003] In the ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12Q1/686C12Q1/04C12N15/11C12R1/645
CPCC12Q1/686C12Q1/6895C12Q2600/166C12Q2563/185
Inventor 徐平田飞唐蜀昆施佳辉周文婧高林瑞高慧英职晓阳丁章贵刘韬
Owner MENGHAI TEA IND
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