Construction of Tobacco Bax lnhibitor-1 ihpRNA Gene Silencing Vector and Transformation into Agrobac

• 作 者: ZHU Wenhua XIE Meng LIN Yuheng YANG Mingyu MA Qiumin TIAN Huiqin QU Guiqin
• 机 构: College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083,
• 出 处: 东北农业大学学报（英文版）
• 发表日期: 2011年2期
• 关键词: Bax inhibitor-1 silencing vector constrution ihpRNA

The plasmid pGSA1285 was first modified by substituting its GUS sequence with the Chalcone synthase intron fragment from vector pFGC5941 to get the plant silencing expression vector that contained Kanamycin resistance site and was named as pGSA2285. Using PCR-based amplification, two different restriction sites at both ends of tobacco Bax inhibitor-1 (NtBI-1) gene were created, respectively, which made the construction of ihpRNA gene silencing vector more efficiently. Then, NtBI-1 genes were inserted into Multiple Cloning Site (MCS) of pGSA2285 respectively to form Bax inhibitor-1 ihpRNA gene silencing vector, named as pGSA4285, containing sense and anti-sense BI-1 sequence which was spliced by chalcone synthase intron. Combined PCR identification and enzyme restriction analyses, the results showed that Bax inhibitor-1 ihpRNA gene silencing vector had been constructed and transferred into Agrobacterium tumefaciens EHA105 successfully, which laid a foundation for the further study on the function of BI-1 in plant PCD regulation.