A kind of Phoebe sesquiterpene synthase sgstps2 and its coding gene and application
A technology of sesquiterpene synthase and eucalyptus, applied in application, genetic engineering, plant genetic improvement and other directions
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[0028] 1. Clone the oleifera sesquiterpene synthase gene SgSTPS2, construct a cloning vector and transform prokaryotic cells
[0029] Extract the RNA of the stem tissue of Phoebe oleifera, and use the reverse transcriptase M-MLV to reverse transcribe the synthesized cDNA. Using the cDNA as a template, the forward primer is: 5'-ATGTCGGTTGCAGCTTTAGC-3', the reverse primer is: 5'-TCATGCGACAGGATTTATGA-3', and PCR amplification is performed using Ex Taq DNA polymerase from TakaRa Company; the PCR conditions are: : 94°C for 5min; 94°C for 30s, 55°C for 1min, 72°C for 1min30s, 35 cycles; 72°C for 10min. The PCR product was detected by 1% agarose gel electrophoresis, and the results were as follows: figure 1 as shown, figure 1 M is the DNA marker DL2000, and the fragment size of the target gene SgSTPS2 is about 1700bp, which is in line with expectations.
[0030] Use the method of agarose gel electrophoresis gel recovery kit to recover the target gene fragment, carry out TA cloning...
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