Comparative study between two construction methods of short hairpin RNA expression vector targeting Epsteinª²Barr virus LMP1 gene

    WANG Yiª²Qin, YANG Yuª²Cheng, HONG Suª²Ling

    Department of Otorhinolaryngologyª²Head and Neck Surgery, First Affiliated Hospital, Chongqing University of Medical Sciences, Chongqing 400016, China

    ¡¾Abstract¡¿ AIM: To explore the differences between two construction methods of short hairpin RNA expression vector targeting Epsteinª²Barr virus(EBV) latent membrane proteinª²1(pshLMP1), so as to look for a more stable and convenient way of constructing the special shRNA vector. METHODS: The siRNA coding sequences targeting EBVª²LMP1 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. ......