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Passion flower reference gene as well as screening method and application thereof

An internal reference gene and screening method technology, applied in the field of molecular biology, can solve problems such as restricting the research process of passionflower cloning, etc., and achieve the effects of shortening detection time, optimizing PCR amplification procedures, and improving detection efficiency.

Active Publication Date: 2020-04-10
GUANGXI ZHUANG AUTONOMOUS REGION ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This seriously restricts the research process of important trait gene mapping and cloning of Passiflora

Method used

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  • Passion flower reference gene as well as screening method and application thereof
  • Passion flower reference gene as well as screening method and application thereof
  • Passion flower reference gene as well as screening method and application thereof

Examples

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Embodiment 1

[0031] A Passiflora internal reference gene, the internal reference gene is C27182, and the nucleotide sequence of the internal reference gene C27182 is shown in SEQ ID NO.1.

Embodiment 2

[0033] A primer for PCR amplification of an internal reference gene C27182, the forward sequence of the nucleotide sequence is shown in SEQ ID NO.2; the reverse sequence is shown in SEQ ID NO.3.

Embodiment 3

[0035] A screening method for Passiflora internal reference gene C21209, comprising the following steps:

[0036] S1. Download 4 sample data from the NCBI database, the sequence accession numbers are: SRX4224487, SRX4224486, SRX4224485 and SRX4224484;

[0037] Use Trinity (version r20131110) software, parameter min_kmer_cov 2 to assemble the sequence;

[0038] Use RSEM (v1.2.3) to analyze the expression level of predicted sequences;

[0039] Functional annotation of gene sequences using BLAST (v2.2.31) and databases NR, COG, KOG, eggNOG, KEGG, GO, Pfam and Swiss-prot;

[0040] Calculate the stability of the expression value of all Passiflora genes in 4 samples, use the coefficient of variation (CV) to evaluate, select the gene with CV value less than 0.2, and cDNA ≥ 1000bp as the candidate memory gene;

[0041] S2. Among the candidate genes, according to the expression value of the Passiflora gene, five genes with moderate expression (FPKM≈25) were selected, namely C24822 (N...

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Abstract

The invention provides a passion flower reference gene as well as a screening method and an application thereof, which belongs to the field of molecular biology. The RNA-Seq sequencing data is used for analyzing stable genes with the medium expression level to serve as the candidate reference genes; five genes with medium expression quantity (FPKM is approximately equal to 25) are selected, primerdesigning is carried out, verifying is carried out in mRNA of different tissue parts of passion flower varieties with obvious phenotypic difference, the expression stability of each candidate reference gene is estimated by utilizing software, and finally the proper passion flower reference gene C27182 is screened out. The C27182 can be stably expressed in stems, leaves, flowers and fruits of Tainung No.1 and golden fruit No.2. The reference is provided for subsequent scientific research related to passion flower gene expression.

Description

technical field [0001] The invention relates to the field of molecular biology, in particular to an internal reference gene of passionflower and its screening method and application. Background technique [0002] Passiflora is an important fruit tree in southern my country. Its fruit has an aromatic smell, is rich in carbohydrates, vitamins and mineral elements, and has high nutritional value and good economic benefits. Passion fruit juice contains 165 kinds of fruit aromas such as pineapple, banana, strawberry, apple, sour plum, mango, etc. . Its natural raw juice is known as the natural nutrient concentrate and queen of tropical fruits for its bright color, unique and strong aromatic flavor, rich nutrition and health care effect on the human body. Passiflora fruit not only has high nutritional value, promotes body fluids and quenches thirst, but also has the effect of disease prevention and fitness. According to the determination, passion fruit juice contains more than ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12Q1/6806C12Q1/686C12N15/11
CPCC12Q1/6806C12Q1/686C12Q1/6895C12Q2600/166C12Q2563/107C12Q2545/114
Inventor 吴艳艳刘洁云田青兰杨行海黄伟华牟海飞
Owner GUANGXI ZHUANG AUTONOMOUS REGION ACAD OF AGRI SCI
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