PMI基因作为选择标记的植物表达载体构建及转化白花丹参体系的建立(4)
[Abstract] Objective： To construct plant expression pCAMBIA1301-PMI by substituting PMI for hygromycin resistance gene in pCAMBIA1301 and obtain transgenic Salvia miltiorrhiza f. alba using PMI-mannose selection system. Method： The 6-phosphomannose isomerase gene (PMI) of Escherichia coli was amplified by PCR. Sequence analysis showed that it shared 100% amino acids identities with the sequences of PMI genes isolates reported in the NCBI. Based on pCAMBIA1305， the plant expression pCAMBIA1305-PMI was constructed successfully by substituting PMI for hygromycin resistance gene in pCAMBIA1305. pCAMBIA1305-PMI was transformed into Agrobacterium tumefaciens LBA4404， and then the leaves of S. miltiorrhiza f. alba were inoculated in LBA4404 with pCAMBIA1305-PMI. Result： Plant expression pCAMBIA1301-PMI was successfully constructed and the leaves of S. miltiorrhiza f. alba inoculated in LBA4404 with pCAMBIA1305-PMI were selected on medium supplemented with a combination of 20 g·L-1 mannose and 10 g·L-1 sucrose as a carbon source. The transformation efficiency rate was 23.7%. Conclusion： Genetic transformation was confirmed by PCR， indicating that a new method for obtaining transgenic S. miltiorrhiza f. alba plants was developed using PMI-mannose selection system.
[Key words] 6-phosphomannose isomerase(PMI)； mannose； Salvia miltiorrhiza f. alba； selection marker of genetic transformation
doi：10.4268/cjcmm20140712, 百拇医药(陶如 张友灿 方茜 史仁玖 李艳玲 黄璐琦 郝岗平)
[Key words] 6-phosphomannose isomerase(PMI)； mannose； Salvia miltiorrhiza f. alba； selection marker of genetic transformation
doi：10.4268/cjcmm20140712, 百拇医药(陶如 张友灿 方茜 史仁玖 李艳玲 黄璐琦 郝岗平)