Picea wilsonii transcription factor PwHAP5 and coding gene and application thereof
A technology for encoding gene and transgenic plant, which is applied to the transcription factor PwHAP5 of P. chinensis and its encoding gene and application field
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0042] Example 1, Discovery of transcription factor PwHAP5 and its coding gene
[0043] 1. Processing of green stem pollen
[0044] On the sunny morning when the inflorescences of Qinggan (Zhang LY, Lin JX, et a1.Plant Science, 2007, 172: 1210-1217.) (Piceawilsonii Mast., collected from the Institute of Botany, Chinese Academy of Sciences, Beijing) were open, use sterilized forceps to take Put the yellow pollen pieces on white clean paper, put them into a sterilized dry glass bottle, dry them in the shade indoors, then pack them into dry glass bottles, seal them, and store them in a -20°C refrigerator spare. The pollen stored in the -20°C refrigerator was transferred to 4°C for recovery for 12 hours, and then transferred to room temperature for 2 hours of recovery. The revived pollen was placed in a liquid medium and germinated at room temperature (25±1°C). The composition of the medium includes: 12% sucrose, 0.03% calcium nitrate or 0.1% calcium nitrate, 0.01% boric acid, ...
Embodiment 2
[0051] Example 2, analysis of tissue expression of PwHAP5 gene by semi-quantitative RT-PCR
[0052] Extract the RNA from the pollen, needle leaf, bark and root tissue of the green stem, and reverse transcribe to synthesize cDNA; The primers were used for PCR amplification to detect the expression of PwHAP5 gene in different tissues of A. sinensis. At the same time, the EF1-α gene was used as an internal reference, and the 5' primer for amplifying the EF1-α gene was: 5'-AACTGGAGAAGGAACCCAAG-3', and the 3' primer was: 5'-AACGACCCAATGGAGGATAC-3'.
[0053] PCR reaction conditions: pre-denaturation at 94°C for 5 minutes; then 10 sec at 94°C, 10 sec at 56°C, 20 sec at 72°C, a total of 28 cycles; extension at 72°C for 5 min.
[0054] The expression of PwHAP5 gene in each tissue is as follows: figure 1 shown. The results showed that PwHAP5 was expressed in all tissues, the expression level was lower in roots and the highest in needles.
Embodiment 3
[0055] Embodiment 3, the acquisition of transgenic Arabidopsis and its detection
[0056] 1. Construction of recombinant expression vector
[0057] 1. Acquisition of PwHAP5 gene
[0058] Prepare the following primer pairs:
[0059] Primer 1: 5'-ATGGATCAGCAGCAGCCCA-3';
[0060] Primer 2: 5'-TCAACTGCTGCCATGAGGAGG-3'.
[0061] With Qinggan (Picea wilsonii Mast.,) (Zhang LY, Lin JX, et al.Plant Science, 2007, 172: 1210-1217.) (the non-patent literature that has recorded this material is Zhang LY, Hao HQ, Lin JX .The localization of Rac GTPase in Picea willsonii pollen tubes implies roles in tube growth and the movement of the tube nucleus and sperm cells.Plant Science, 2007, 172:1210-1217, the public can obtain from Beijing Forestry University) cDNA as a template, using The above primers 1 and 2 were amplified by PCR. Sequencing showed that the nucleotide sequence of the amplified product was shown in sequence 1 in the sequence listing. There were 606 nucleotides in sequence 1...
PUM

Abstract
Description
Claims
Application Information

- R&D
- Intellectual Property
- Life Sciences
- Materials
- Tech Scout
- Unparalleled Data Quality
- Higher Quality Content
- 60% Fewer Hallucinations
Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.
© 2025 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com