Studies On The DREB1A Cloning And Correlation Analysis Of Populus Simonii

In this thesis taking the Populus simonii as the experiment material which were propagated according to their morphological characters by using the different drought stress and water deficit to treat with them. We extracted total RNA from plant leavies, then countertranscripted total RNA into complement DNA, amplified with the primer specific to DREBIA via PCR, detected the PCR products via Agarose gel electrophorecis and acquired a PCR fragment with the size of more than 800 base pairs. The product was purified, recovered and ligated to PGEM-T easy vector. Transformed into competence cell-DH5a Escherichia coli. Then screened the recombinant with a-complement experiment. The recombination ratio was up to 93%. Picked up some blue and white colony respectively, extracted with plasmid DNA and diluted to the one hundred times than it. The diluted plasmid DNA was amplified as PCR template. The results were shown that only the white colony can be amplified and detect a PCR product with the similar size to prior PCR product. By sequencing the colony included a complete open reading frame which encodes with polypeptide comprised of 251 amino acids. The sequence of PCR product was compared with Genbank database online. The result of alignment was that there are 87% homology with between PCR product and DREBIA of Arabidopsis thaliana. Based on this, it was followed that DREBIA correlated with drought resistance had been isolated from populus simonii, which provided with the invaluable experience, as well as, a new research path was groped for improving tree drought resistence to stress.