M yb28基因表达载体的构建及拟南芥转化

Construction of Expression Vectors of Brassica chinensis M yb28 Gene and Transformation to Arabidopsis thaliana

利用通过RT-PCR扩增到的M yb28(GenBank注册号:HQ270468)基因分别构建正义和反义植物表达载体,采用冻融法转入农杆菌LBA4404菌株,通过花序浸泡法对Myb28基因缺失的拟南芥进行了遗传转化,经RT-PCR和酶切鉴定,结果表明Myb28正义和反义真核表达载体构建成功,经基因组PCR鉴定表明正义表达载体已成功整合到拟南芥基因组中。

A full-length cDNA encoding Myb28 from Brassica chinensis was isolated using RT-PCR.The recombinant plasmids were transferred into Agrobacterium tunefacien LBA4404 by freeze-thaw method,and then were used to infect mutant Arabidopsis thaliana.Using PCR,we proved that the sense and antisense expression vector were constructed successfully and the genes were integrated into Arabidopsis thaliana.