EB virus LMP1 gene-targeted shRNA expression vector Comparative Study.docVIP

 PAGE \* MERGEFORMAT 13 EB virus LMP1 gene-targeted shRNA expression vector Comparative Study [Keywords:] Herpesvirus 4, human; RNA interference; genetic vectors Comparative study between two construction methods of short hairpin RNA expression vector targeting EpsteinBarr virus LMP1 gene [Abstract] AIM: To explore the differences between two construction methods of short hairpin RNA expression vector targeting EpsteinBarr virus (EBV) latent membrane protein1 (pshLMP1), so as to look for a more stable and convenient way of constructing the special shRNA vector. METHODS: The siRNA coding sequences targeting EBVLMP1 were designed; pshLMP1 expression vector was achieved by the conventional annealing method or PCR method, and the differences of constructive strategies, construction processes and sequencing results between the two methods were analyzed. RESULTS: pshLMP1 expression vector was obtained by both methods but better cloning efficiency was achieved by PCR method with rare base deletion and mutation. CONCLUSION: PCR method is a more efficient way to construct the more stable pshLMP1 expression vector. [Keywords] herpesvirus 4, human; RNA interference; genetic vectors [Abstract] Objective: To explore the two kinds of EB virus LMP1 gene construct shRNA expression vector (pshLMP1) method is different from looking for a more stable and convenient shRNA expression vector targeted ways. Methods: The design for the EB virus LMP1 gene encoding specific siRNA sequences, respectively, a traditional double-strand annealing method and the novel double-stranded PCR method to build pshLMP1, compare the two methods in the design principles, construction methods and identification of results on the difference. Results: The two methods can be pshLMP1 recombinant plasmid, but the Construction of double-stranded PCR method is efficient and easy to cause base deletions and mutations. Conclusion: Construction of EB viral double-stranded PCR method than the traditional do