PDZ1 - 2 domains and recombinant adenovirus shuttle vector construction.doc

 PAGE \* MERGEFORMAT 14 PDZ1 / 2 domains and recombinant adenovirus shuttle vector construction Study: High-Fang, Hu book group, Sun Dong, Zhang Guangyi [Abstract] Objective To construct the domain of PSD-95 PDZ1 / 2 recombinant adenovirus. Ways to different guanidine sulfate cyanide - phenol - chloroform extraction step of total RNA as template, obtained by RT-PCR PDZ1 / 2 of the cDNA, with adenovirus shuttle vector with T4 DNA ligase, ligation products transformed into E. coli JM109 were screened, and sequenced. Results obtained by RT-PCR PDZ1 / 2 of the cDNA, enzyme digestion can be observed to PDZ1 / 2 and pAdTrack -CMV two bands, PDZ1 / 2 sequencing patterns consistent with those reported. Conclusion The target gene construct containing PDZ1 / 2 of the recombinant adenovirus shuttle vector. [Keywords:] PDZ1 / 2, RT-PCR, adenovirus shuttle vector, sequencing Abstract: Objective To construct recombinant pAdTrack-CMV-PDZ1 / 2. Methods The total RNA extracted with single-step method by acid guanidinium thiocyanate-phenol-chloroform extraction was used as the template, and the cDNA of PDZ1 / 2 domain was amplified by RT-PCR; the former was subsequently cloned into the shuttle vector pAdTrack-CMV (CLONTECH) by ligation of T4 DNA ligase. Thereafter, the sequence was transfected into E. coli Electro-Cell JM109 for screening and determination.Results The cDNA of PDZ1 / 2 was amplified by RT-PCR. The recombinant contained two bands of pAdTrack-CMV and PDZ1 / 2 by restriction enzyme digestion. The sequence chromatogram of cloned PDZ1 / 2 was in accordance with GenBank Database. Conclusion Recombinant of the targeted pAdTrack-CMV-PDZ1 / 2 is successfully constructed. Keywords:: PDZ1 / 2; RT-PCR; pAdTrack-CMV; sequence determination Currently, ischemic stroke on neuronal damage and death, there are three kinds of molecular mechanisms of the hypothesis: glutamate excitotoxicity, calcium overload and free radicals. Among them, the glutamate recepto