Fast-growing aquatic plant nitrate transporter genrt2.1 and its coding gene and application
A technology that encodes genes and nitrates, which is applied in the direction of plant genetic improvement, application, microorganisms, etc., and can solve problems such as inability to transport or grow
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0061] Embodiment 1, the acquisition of GeNRT2.1
[0062] 1. Discovery of GeNRT2.1
[0063] 1. Total RNA extraction and cDNA synthesis of dwarf pearls
[0064] Using the RNAprep Pure Plant kit from TIANGEN Company, the total RNA of dwarf pearls was extracted according to the kit instructions, and cDNA was obtained by reverse transcription.
[0065] 2. PCR amplification
[0066] Using the cDNA obtained by reverse transcription in step 1 as a template, PCR amplification was performed using degenerate primers H5 and H3 to obtain a PCR product. The primer sequences are as follows: H5: GGNGCNGAYCARTTYGAYG; H3: AVYTCYTCNACYTGNGTNAC. The PCR amplification program was: 95°C pre-5min; 94°C denaturation 30s; 52°C annealing 30s; 72°C extension 1min, 35cycle; 72°C, 10min.
[0067] 3. Electrophoresis and sequencing
[0068] After the amplification was finished, 1% agarose gel electrophoresis was detected, and the PCR product was connected to the T carrier, and the sequencing results s...
Embodiment 2
[0126] Embodiment 2, verification of GeNRT2.1 gene function
[0127] In this example, the function of the GeNRT2.1 gene was verified by using the Δynt-Leu double mutant Hansenula polymorpha.
[0128] 1. Construction of transgenic GeNRT2.1 yeast
[0129] 1. Construction of the shuttle plasmid
[0130] Insert the DNA molecule shown in Sequence 4 in the sequence listing between the speI and salI restriction sites of the pYNR-EX vector, and keep other sequences of the pYNR-EX vector unchanged to obtain the recombinant vector pYNR-GeNRT2.1.
[0131] 2. Linearization
[0132] pYNR-GeNRT2.1 was digested with BstpI and linearized to obtain linearized pYNR-GeNRT2.1, which was subjected to 1% agarose gel electrophoresis ( Figure 4 ), recover and purify the target product, use a spectrophotometer for quantification, and set aside.
[0133] 3. Preparation of double mutant Hansenula polymorpha (△ynt-Leu) competent cells
[0134] ①Pick a single colony of △ynt-Leu double mutant Hansenu...
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More 
PatSnap Eureka turns technology decisions into work you can execute. Powered by our Innovation Knowledge Graph, it runs expert workflows across engineering, life sciences, materials and intellectual property. Get your review-ready output in minutes.
