Artificial Chimeric Promoters Construction And Stress Responsive Activity Analysis

Abiotic stresses such as salinity, drought and low temperature affect plant growth and cropproduction extensively. Using gene engineering method to cultivate resistant plant is the importantapproach for resistant biology research. The regulation of gene expression plays an important rolein plant in response to stresses. There are many stress-related genes in plant. Plant can regulatesstress-related genes expression and improve the tolerance under environmental stresses. Now somany stress-related genes have been cloned and used for the resistance-engineering research, andsome transgenic plants have been got which could tolerate the stresses to some extent. The mostfrequently used promoter for regulating the interested genes expression is CaMV35S promoterwhich can overexpress the interested genes, but the expression is neither environment, tissue nortime specific, it is constitutive. Though it can improve the plant tolerance using the CaMV35Spromoter to express the related genes in stress environment, the expression of resistant proteinwastes the energy in normal environment and the accumulation of the protein can inhibit the plantsnormal metabolism, causing the morphology change of the plant, resulting in growth anddevelopment retardment of transgenic plants. In order to solve the problem as above, people canuse the stress specific induced promoters, but the activity of these promoters identified as far stillcan't content the need of related research. So the experiment attempts to construct artificialchimeric promoters which can be strongly induced by several stresses.Now so many stress induced genes and their promoters have been isolated, and some promoterswere identified in detail, confirmed the key fragments by which the promoters performed theirfunction mainly. Scientists have also indentified a series of cis-elements which interacted with thetranscription factor,such as drought-responsive element DRE,ABA-responsive element ABRE,light-responsive element G-box and so on. We got five stress-induced genes'promotersequence(eg. rd29A,erd1,cor15a,lti30,kin1)on the basis of previous publications, and thesepromoters are all induced by several stresses. They all have stress-responsive elements and the keyfragments have been confirmed. We got the key fragments from the five promoters using PCRmethod. Because rd29A promoter is relatively strongly induced one, on the basis of 269bp...