ZmLAX3 protein, and coding gene and application thereof
A protein and coding technology, applied in the field of ZmLAX3 protein and its coding gene and application, can solve the problems of less QTL and increased versatility of molecular marker-assisted selection
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Embodiment 1
[0078] Embodiment 1, ZmLAX3 gene cloning
[0079] 1. Extraction and purification of corn RNA
[0080] Utilize the Trizol method to extract the total RNA of corn inbred line Qi 319 (hereinafter also referred to as wild-type corn), and the specific method steps are as follows:
[0081] Take a DEPC-treated 1.5ml centrifuge tube, add 1ml Trizol; take 100mg of corn material, place it in liquid nitrogen and grind it repeatedly to powder, transfer it to the above centrifuge tube, mix well, and let stand at room temperature for 5 minutes; 12000rpm, 4°C , centrifuge for 10 minutes, transfer the supernatant to a new centrifuge tube, add 200 μl chloroform, shake vigorously for 15 seconds, let stand at room temperature for 3 minutes; centrifuge at 12000 rpm, 4°C for 15 minutes, take the supernatant to a new centrifuge tube ;Add 250μl isopropanol and 250μl high salt solution, mix by inverting, and let stand at room temperature for 10 minutes; 12000rpm, 4℃, centrifuge for 10 minutes, pour ...
Embodiment 2
[0093] Example 2, Functional Research of ZmLAX3 Gene
[0094] 1. Construction of plant recombinant vector pZP211 UBI::ZmLAX3
[0095] The RNA of maize inbred line Qi 319 was extracted, reverse-transcribed to obtain cDNA as a template, and PCR was amplified with upstream primers and downstream primers to obtain a 1897bp PCR amplification product of ZmLAX3 gene.
[0096] Restriction endonucleases BamHI and Sac Ⅰ double-digest the ZmLAX3 gene PCR amplification product, and BamHI and Sac Ⅰ double-digest the pZP211::UBI expression vector "Cloning of ZmAATP Gene and Construction of Genetic Transformation Vector in Maize", Shandong Journal of Agricultural University (Natural Science Edition), 2012, 43(3) 321-327; the public can obtain it from Shandong Agricultural University), and the operation steps are carried out according to the instructions of T4 DNA ligase of Fermentas company. Then the ligation product was transformed into Escherichia coli DH5α competent cells, and cultured o...
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