Cotton cell wall extensin gene GbEXPATR and application
A technology of extensin and cell wall, which is applied in cotton cell wall extensin gene GbEXPATR and its application field, can solve the problems of unclear biological function and achieve the effect of promoting elongation and high accuracy
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0033] Example 1: GbEXPATR gene isolation clone and its expression characteristics in different tissues of cotton
[0034] In the preliminary work of the applicant's State Key Laboratory of Crop Genetic Improvement, the cDNA library of sea-island cotton (Tu, L.L.; et al. Genes expression analyzes of sea-island cotton (Gossypium barbadense L.) during fiber development. Plant Cell reports2007,26(8),1309-1320.) isolated a cDNA that is highly homologous to the upland cotton cell wall expansin gene (gene accession number: AF043284) but lacks the second domain (see SEQ ID NO: 1), bioinformatics analysis showed that the cDNA contained a complete ORF (see SEQ ID NO: 3), and it was predicted by bioinformatics that this ORF could translate an incomplete α-expansin protein (the coded The amino acid sequence of the protein is shown in SEQ ID NO: 2), and we named this fragment GbEXPATR gene. The gene is highly expressed at various stages of fiber elongation, and 10DPA fiber total RNA is e...
Embodiment 2
[0035] Embodiment 2: Construction and transformation of overexpression vector
[0036] In order to verify the function of the GbEXPATR gene in cotton, the applicant constructed an overexpression vector to transform cotton.
[0037] According to the ORF region of the GbEXPATR cDNA obtained in Example 1 (see the region corresponding to 52-306 bases in SEQ ID NO: 1), the overexpression primer pair was designed (the DNA sequence of the primer pair is described in this paragraph), we are in NcoI and BstEII restriction sites and protective bases were added to the two ends of the primer pair, respectively, and the primer pair was named as primer GbEXPATRPoef and GbEXPATRPoer, and then the cDNA of the GbEXPATR gene was used as a template for PCR amplification, and the obtained PCR product was passed The method of digestion and ligation was connected to pCAMBIA2301 digested with the same enzyme m on the vector (pCAMBIA2301 m The precursor of the vector is pCAMBIA2301 donated by the C...
Embodiment 3
[0070] Example 3: Correlation Analysis of GbEXPATR Transgenic Lines
[0071] 1. Molecular identification of GbEXPATR transgenic lines
[0072] Three transgenic families were obtained through the overexpression of GbEXPATR gene, which were named as OE2, OE3 and OE4. Genomic DNA was extracted from the obtained transgenic families (according to conventional methods), and analyzed by Southern blot to identify the effect and copy number of the transgene. Genomic DNA extraction methods and Southern experiments of transgenic families refer to the methods reported by J. Sambrook et al., translated by Huang Peitang et al., Molecular Cloning Experiment Guide (Third Edition), Science Press, 2002 edition. see results Figure 4 shown.
[0073] 2. Expression analysis of GbEXPATR transgenic lines
[0074] Extract the total RNA of the transgenic line by single plant (J. Sambrook et al., Huang Peitang et al. translation, Molecular Cloning Experiment Guide (Third Edition), Science Press, 20...
PUM
Property | Measurement | Unit |
---|---|---|
length | aaaaa | aaaaa |
Abstract
Description
Claims
Application Information
- R&D Engineer
- R&D Manager
- IP Professional
- Industry Leading Data Capabilities
- Powerful AI technology
- Patent DNA Extraction
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