Plant drought resistance and salt tolerance associated protein EeABF6, coding gene and application thereof
A related protein and salt-tolerant technology, applied in the field of genetic engineering, can solve the problems of inability to achieve plant stress resistance and small improvement
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Embodiment 1
[0087] Example 1: cDNA cloning of EeABF6 gene related to drought resistance and salt tolerance of E. elongatum.
[0088] The 45-day-old E. elongatum seedlings were subjected to drought treatment for 5 hours, and Trizol was used to extract the total RNA of E. elongatum. 5'RACE kit (5'RACE System for Rapid Amplification of cDNA Ends Kit) (GIBCOBRL, CAT.NO.18374-058) and 3'RACE kit (3'RACE System for Rapid Amplification of cDNAEnds Kit) (GIBCOBRL, CAT .NO.18373-019) to obtain the EeABF6 gene.
[0089] Total RNA was extracted from Echinopsis elongatum seedlings with Trizol, and cDNA was obtained by reverse transcription with superscript II (invitrogen) reverse transcriptase. Primers P1 and P2 were designed according to the sequence of the coding region of EeABF6 gene. Using the cDNA obtained by reverse transcription as a template, PCR amplification was performed with primers P1 and P2. The sequences of primers P1 and P2 are as follows:
[0090] P1: 5'-GCGGGATGGACTTTCCGG-3',
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Embodiment 2
[0094] Embodiment 2: Cultivate drought-resistant, salt-tolerant transgenic plants with EeABF6 gene
[0095] 1. Construction of recombinant expression vector
[0096] 1) Construction of 35S-EeABF6 recombinant expression vector
[0097] Use the cDNA obtained by reverse transcription of the total RNA of Echinopsis elongatum as a template, and perform PCR amplification with specific primers containing SmaI and XbaI adapter sequences; then SmaI and XbaI double-enzyme digest the PCR product, recover it, and forward Insert the vector pBI121 between the SmaI and XbaI restriction sites behind the CaMV 35S promoter to obtain the recombinant vector 35S-EeABF6.
[0098] The primer sequences are as follows:
[0099] EeABF6[SmaI]5'-GCGCCCGGGGCGGGATGGACTTTCCGG-3'
[0100] EeABF6[XbaI]5'-TGCTCTAGACCAAGGGCCCGTCAGCGT-3'
[0101] 2) Construction of 29A-EeABF6 recombinant expression vector
[0102] Use the cDNA obtained by reverse transcription of the total RNA of Echinopsis elongatum as a t...
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