Studies on the Construction of cDNA Library in Drought-Induced Popμlus Simonii and Preparation of Library-Screening Probe with the mRNA Differential Display
|School||Shanxi Agricultural University|
|Keywords||Drought induced P. simonii cDNA library Differential Display Recover|
To improve and enhance the drought resistance of plants is an important part of the study of plant stress resistance . With the development of molecular biology techniques , drought resistance of plants through genetic engineering to improve the majority of scientists believe is a quick and effective way . As upstream technology of genetic engineering - cloning have developed rapidly in the nineties of the last century , a brief overview of the plant gene cloning in recent years new technologies, new methods , with emphasis on the differential display (DDRT-PCR) described . In this study, lobular Yang (Popμlus simonii) as experimental materials to build a drought -induced cDNA library of lobular Yang . And by differential display technology to probe for screening of the cDNA library , made ??the following findings : ZAP phage vector system , and built the original storage capacity of 2 × 10 5 sup> pfu / ml drought treatment sample library and 0.8 × 10 5 sup> pfu / ml of the control libraries. After amplified library drought treatment sample library increased to 2.0 × 10 10 < / sup> pfu / ml, and the control library increased 1.2 × 10 8 < / sup> pfu / ml . DDRT-PCR results demonstrated by silver staining drought treatment samples and controls a total of nearly 5,000 bands appeared , drought -like with still greater differences between the differences were picked with 35 . Recovery of the difference with the secondary amplification and direct sequencing EST BLAST comparison , cDNA fragments were found associated with drought . The study also improved recovery of the bands on polyacrylamide gel electrophoresis , has made a good recovery results . Next library screening , gene get to do the necessary preparations .