烟草新驱动蛋白NtTKR过表达对酵母生长的影响

为了解烟草新驱动蛋白NtTKR功能信息,构建了EGFP融合的NtTKR基因酵母表达载体并转入酿酒酵母W303诱导表达,荧光显微镜观察结果表明,诱导条件下的转化细胞发出明亮的绿色荧光,表明转入的NtTKR得到了高效表达.转化株在诱导和非诱导条件下培养3d,可见NtTKR过表达菌落比对照小的多;但通过绘制8h内生长曲线发现二者细胞数目无显著差别;表明NtTKR过表达对酵母细胞生长的抑制可能更多的是因为抑制其体积增大,而非分裂速度减慢引起的细胞数目降低.已知NtTKR在烟草受精卵、发育各个时期的胚、幼叶、根尖及茎尖等细胞分裂旺盛部位均优势表达,推测该基因在烟草中可能与上述各部位新生细胞的体积增大相关.

Effect of Over Expression of Tobacco new Kinesin NtTKR on the Growth of Yeast

In order to know the function of new kinesin NtTKR in tobacco,NtTKR and EGFP fusion fragment was inserted into yeast expression vector pYES2;the transformed W303 strain was then induced by galactose and raffinose containing medium;under fluorescence microscopy,bright EGFP fluorescence was observed from the transformed cells which was induced.It suggests that the transformed NtTKR was expressed efficiently.After growing on plate for 3days,more smaller colonies were found on induced plate.But from the growth curve in 8hours,no dramatically difference was found between induced transformed cells and control.The results show that the inhibition of over expression of NtTKR to the growth of yeast cells is mainly caused by the inhibition of cell enlargement,but not the reduction of cell number.NtTKR is predominant expression in every development stage embryo,young leaves,root tip and stem tip and other area of tobacco where cell division takes place at a rapid rate,it might involve in the new cell enlargement in all areas mentioned above.