The Establishment Of Competence Cell Transformation System And The Construction Of Transient Expression Vector

This study was carring out in the lab of vegetable germplasm resource and biotechnology in the horticultural college of the Northwest A&F university. In this study,we designed primer and cloned the induction promoter rd29A from DNA of Arabidopsis thaliana.After sequencing the promoter, we analysis its sequence. And then we constructed transient expression vector using rd29A and PBI221.The main results were as follow:1 The influence of different prepatations on the transformation efficiency of E.coli competent cells was studied. The results showed that the efficiency of super E.coli competent cells was higher than common one. And the efficiency of super E.coli competent cells increased 550% over the common one. Meanwhile the influence of different transformation method was studied. A fast plasmid transformation system was established.2 In this study,we designed primer and cloned the induction promoter rd29A from DNA of Arabidopsis thaliana.Then we checked its sequence in the GenBank.After analysising,we found that the induction promoter rd29A contains TATA-box between 370 and 375 base pair,CAAT-box between 302 and 305 base pair,drought response element DRE between 556 and 565 base pair,ABA response element ABRE between 719 and 724 base pair.3 In this study we constructed induction transient expression vector using rd29A and PBI221.firstly we analysised the enzyme cutting sites of induction promoter rd29A and vector PBI221.Then we dealed the promoter rd29A and vector PBI221 using restriction enzyme HindIII and XbaI.The products were linked by T4 DNA ligase under 16℃overnight.In the end we transfered the product into competence cell.After cultivation we can get the clone of PBI221-rd29A.We checked it with the method of enzyme cutting,and the result showed we successfully constructed the induction transient expression vector PBI221-rd29A.