利用农杆菌系统将超甜定基因导入烟草

Introduction of Thaumatin Gene into Tobacco through Agrobacterium -mediated System

由ｐＵＲ５２８构建中间载体ｐＥＨＴ９，再构建超甜定植物表达载体ｐＢＩＴ７。通过直接转化法将ｐＢＩＴ７导入农杆菌，然后利用农杆菌系统转化烟草。经ＰＣＲ、ＰＣＲ－Ｓｏｕｔｈｅｒｎ和Ｓｏｕｔｈｅｒｎ杂交证实，超甜定基因已整合到烟草基因组中。

pEHT9 was constructed from pUR528, and used for the construction of plant thaumatin expression vector pBIT7. Then pBIT7 was introduced to Agrobacterium through direct transformation. Subsequently tobacco was transformed through Agrobacterium -mediated system. It has been confirmed that thaumatin gene has integrated into the genome of transgenic tobacco by PCR, PCR-Southern and Southern blot analyses.