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

A technology of internal reference genes and screening methods, 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, improving reliability, and optimizing PCR amplification procedures

Active Publication Date: 2020-04-03
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 PeNADP as well as screening method and application thereof
  • Passion flower reference gene PeNADP as well as screening method and application thereof
  • Passion flower reference gene PeNADP as well as screening method and application thereof

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Experimental program
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Effect test

Embodiment 1

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

Embodiment 2

[0033] A primer for PCR amplification of the internal reference gene PeNADP, nucleotide sequence: the forward 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 PeNADP, 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] The stability of the expression value of the PeNADP gene in the four samples was calculated, and the coefficient of variation (CV) was used for evaluation. The genes with a CV value less than 0.2 and cDNA ≥ 1000 bp were selected as candidate memory genes.

[0041] S2. Among the candidate genes, according to the expression value of PeNADP, 2 genes with the highest expression and 2 genes with medium and ...

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Abstract

The invention provides a passion flower reference gene PeNADP and a screening method and an application thereof, which belong to the field of molecular biology. The RNA-Seq sequencing data is used foranalyzing moderate or highly stable genes in the expression quantity to serve as candidate reference genes; according to the expression quantity value of the PeNADP, two genes with the highest expression quantity and two medium-high expression genes are selected, the primer is designed, verification is carried out on mRNA of different tissue parts of the passion flower variety with obvious phenotypic difference, the expression stability of each candidate reference gene is evaluated by utilizing software, and finally the proper passion flower reference gene PeNADP is screened out. The reference gene PeNADP can be stably expressed in stems, leaves, flowers and fruits of Tainung No.1 and gold berry No.2., and reference is provided for subsequent scientific research related to passion flowergene expression.

Description

technical field [0001] The invention relates to the field of molecular biology, in particular to a Passiflora internal reference gene PeNADP and a screening method and application thereof. 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...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C12N15/11C07K14/415C12Q1/6895G16B25/20G16B30/20
CPCC07K14/415C12Q1/6895C12Q2600/166G16B25/20G16B30/20
Inventor 吴艳艳杨行海黄伟华刘洁云田青兰牟海飞
Owner GUANGXI ZHUANG AUTONOMOUS REGION ACAD OF AGRI SCI
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