Caragana korshinskii transcription factor ERF and gene thereof
A transcription factor and caragana technology, applied in genetic engineering, plant genetic improvement, angiosperms/flowering plants, etc., can solve problems such as unsatisfactory stress-resistant varieties and limited genetic resources related to stress resistance
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Embodiment 1
[0009] Example 1. Construction of a full-length cDNA library of twisted leaves:
[0010] Caragana korshinskii plants are cultivated in a culture bowl, cultivated in a greenhouse for 60 days, and subjected to drought treatment until the whole plant appears wilting. The young leaves are quickly frozen and ground in liquid nitrogen. The total RNA is extracted by TRIzole and dissolved in water. The mRNA is obtained by processing with the mRNA purification kit. Take mRNA for reverse transcription reaction, and then synthesize the second strand. After homogenization, connect with pDNR-LIB vector (see Creator for specific steps) TM SMART TM cDNA Library Construction Kit (Clontech Cat. No. 634903).
Embodiment 2
[0011] Example 2. Random EST sequencing of cDNA library:
[0012] Randomly select 3000 single clones from the library for 5'-end EST sequencing, then BLAST the sequenced sequences, and select ESTs similar to the verified drought-resistant genes.
Embodiment 3
[0013] Example 3 ESTs related to drought resistance by BLAST were selected, and their corresponding clones were tested to obtain the full-length DNA sequence of the candidate stress resistance gene.
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