摘要
采用Trizol法从12℃处理48h的10d龄薄荷叶片中提取总RNA,通过RT-PCR方法扩增获得薄荷醇合成关键酶基因cDNA(命名为LT,ST),连接到pMD18-T载体上,命名为pMD18-T-LT,pMD18-T-ST。结果表明:扩增的薄荷醇合成酶片段全长约1125bp和942bp。将获得的基因成功构建到植物表达载体pCAMBIA1301上,经转化根癌农杆菌GV3101,经菌落PCR进一步鉴定转化结果。实验结果为进一步从分子水平挖掘并利用薄荷醇合成酶关键基因奠定基础。
The method of Trizol was extracted to the total RNA from 10th day peppermint leaf which was processed about 48h under 12℃ RT-PCR was employed to clone cDNA fragment of menthol synzyme gene（ named for LT, ST）.The purpose fragments named pMD18-T-LT.pMD18-T-ST which were connected on the pMD18-T vector. The result indicated： Sequence length of LT and ST were about 1125bp and 942bp. The coloned genes were introduced into the plant expression vector pCAMBIA1301 and transformed into Agrobacterium tumefaciens GV3101.Colony PCR was employed to evaluate transformation result. The study provided material for further exploring menthol synzyme gene in molecular level.
出处
《食品工业科技》
CAS
CSCD
北大核心
2012年第8期239-241,248,共4页
Science and Technology of Food Industry
基金
国家自然基金(31060148)
