Trxf1基因表达载体的构建及在加工番茄中的遗传转化

Construction of Trxf1 Gene Plant Expression Vector and Transgenic in Processed Tomato

Trxf1是硫氧还蛋白家族中的一员,是组成硫氧还蛋白系统的重要组成部分,在植物的氧化逆境中发挥作用。为进一步研究该基因功能,通过RT-PCR方法从加工番茄叶片中克隆Trxf1基因的编码区,成功构建植物表达载体pBI121-Trxf1,采用农杆菌介导法转化番茄,经PCR、RT-PCR分析表明,植物表达载体已成功整合到番茄基因组中。实时荧光定量PCR研究表明,加工番茄Trxf1基因在其根、茎、叶、花等器官中均有表达,且在叶片中表达量最高。

Trxf1 is a member of Thioredoxin family, and is an important part of Thioredoxin system, it takes effect in oxidative stress in plants. In order to study the function of the gene further, we cloned the ORF of Trxf1 gene from tomato leaves by PCR method and successfully constructed the plant expression vector pBI121-Trxf1 . Tomato plants were transformed via Agrobacterium tumef aciens, PCR and RT-PCR analysis indicated that the foreign gene Trxf1 had been integrated into the genome of tomato and had acquired expression. Real-time quantitative PCR showed that the Trxf1 gene were expressed in roots, stems, leaves and flowers from the processed tomato. The expression level was higher than that in other organs.