Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Internal reference genes stably expressed by Lygus green bug under different factors and their application

A technology of internal reference gene and stable expression, which is used in the determination/inspection of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc., and can solve problems such as wrong biological conclusions and wrong interpretation of quantitative data.

Active Publication Date: 2022-05-10
HUBEI UNIV
View PDF3 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, a large number of studies have shown that the mRNA expression level of any internal reference gene is constant under all experimental conditions.
Use of unstable endogenous reference genes as internal references for data normalization can lead to misinterpretation of quantitative data and ultimately lead to erroneous biological conclusions

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Internal reference genes stably expressed by Lygus green bug under different factors and their application
  • Internal reference genes stably expressed by Lygus green bug under different factors and their application
  • Internal reference genes stably expressed by Lygus green bug under different factors and their application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1: Acquisition of Candidate Internal Reference Gene Sequences of Lygus viridans

[0036] The method for obtaining a candidate internal reference gene sequence of Lygus chlorophylla comprises the following steps:

[0037](1) Breeding of Lygus spp.: The population of Lygus spp. comes from the indoor population of the Institute of Plant Protection, Chinese Academy of Agricultural Sciences. It is fed with fresh corn (Zea mays) and green beans (Phaseolus vulgaris), and the breeding temperature is 25-28°C , the relative humidity is 60%-70%, and the light is 16:8 (L:D).

[0038] (2) Obtain candidate internal reference gene sequences: firstly, extract the total RNA of Lygus chrysalis, construct the transcriptome database, and obtain the transcriptome sequence information of the whole chrysalis by Illumina sequencing. Then, based on the above sequence information, 8 housekeeping genes EF1γ (SEQ ID NO: 1), RPL32 (SEQ ID NO: 2), RPL27 (SEQ ID NO: 3), SDH (SEQ ID NO: 4), ...

Embodiment 2

[0039] Example 2: Screening of internal reference genes stably expressed by Lygus aeruginosa under different conditions

[0040] (1) Handling and collection of Lygus green bugs under different factors

[0041] ①Collection of Lygus japonica at different developmental stages: 50 first-instar nymphs, 30 second-instar nymphs, 15 third-instar nymphs, 6 fourth-instar nymphs, 6 fifth-instar nymphs, and 5 immature (1-day age) female adults, 5 sexually immature (1-day-old) male adults, 5 sexually mature (12-day-old) female adults, and 5 sexually mature (12-day-old) male adults, placed in liquid nitrogen for quick freezing, Store at -80°C for later use. Each treatment has 3 biological replicates.

[0042] ②Collection of different tissues of Lygus japonica: Collect the 5th instar nymphs of Lygus japonica, dissect and collect the head, thorax, midgut and residual body in pre-cooled PBS, dissect at least 25 test worms for each sample, and dissect the tissues immediately After quick-free...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

PropertyMeasurementUnit
PCR efficiencyaaaaaaaaaa
Login to View More

Abstract

The invention proposes an internal reference gene combination stably expressed by Lygus viridans under different factors and its application. The specific combination scheme is as follows: RPL27 and RPL32 are stably expressed internal reference genes at different developmental stages, RPL27 and RPL32 are internal reference gene combinations in different tissues, and RPL27 and EF1γ are internal reference gene combinations under dsRNA treatment conditions. Different primer sequences used when the stably expressed internal reference gene is used for fluorescence quantitative analysis are also proposed. The present invention determines for the first time the most stable internal reference gene combination of the green Lygus under the conditions of three factors, and provides a basis for studying the analysis of gene expression levels of the green Lygus at different developmental stages, different tissues, and dsRNA-mediated RNA interference conditions. The selection of suitable internal reference genes has established a solid foundation for the study of the gene function of Lygus chinensis and the development and safety evaluation of RNAi-mediated transgenic crops.

Description

technical field [0001] The invention belongs to the technical field of expression stability of real-time fluorescent quantitative PCR internal reference genes in insect molecular biology, and more specifically, relates to the combination of internal reference genes stably expressed under different factors of the green Lygus bug and its application; Internal reference genes stably expressed in different stages, different tissues, and dsRNA-mediated RNA interference conditions and their applications. Background technique [0002] Apolygus lucorum belongs to Hemiptera, Miridae, and is one of the main pests in cotton planting areas in my country. Its host range is very wide, mainly including cotton, corn, soybean, alfalfa, cruciferous vegetables, tea tree, Flowers and various fruit trees have caused huge economic losses to my country's agriculture and forestry. At present, its prevention and control is mainly based on chemical control, but the long-term use of chemical agents wi...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6888C12Q1/686C12N15/11
CPCC12Q1/6888C12Q1/686C12Q2600/166C12Q2561/113C12Q2563/107C12Q2545/114
Inventor 罗静王奥丽成艳霞郭丽彬徐乐天
Owner HUBEI UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com