荧光素酶作为可视标记整株检测转基因玉米

利用活体成像系统检测转基因玉米中荧光素酶信号整株实时筛选高表达的转基因玉米。结果表明,按照信噪比大于2为高表达转基因阳性植株的标准,获得的T1代66株转pMD35LUC的植株中,11株有荧光信号,其中2株信噪比值大于2;获得的T1代68株转化pMDUBLUC的植株中,30株有荧光信号,6株信噪比值大于2。信噪比值大于2的植株移栽成活7株,经PCR检测均为阳性。2株T1植株的T2代植株无荧光信号值;5株T1植株的T2代植株都有数量不等的植株有荧光信号,其中4株T1株都有信噪比值大于2的T2代植株,经PCR检测均为阳性。本研究跳过一系列大量的操作复杂、耗时、高成本的体外分子检测,直观、准确、实时、低成本地淘汰大量非转基因和表达不强的植株或株系,为玉米转基因植株的筛选提供了有效方法。

Firefly Luciferase as a Vital Marker to Detect Intact Transgenic Maize

Luciferase gene expression in intact transgenic maize plants was detected by in vivo imaging system in real-time.Among 66 T1 plants transferred with pMD35LUC,11 plants were detected with signal and 2 plants with the signal-to-noise ratio(SNR)more than 2.Among 68 T1 plants transferred with pMDUBLUC,30 plants were detected with signal and 6 plants with SNR more than 2.Seven plants of T1 plants with SNR more than 2 were transplanted alive and positively transgenic plants by PCR.The signals were not detected on T2 plants from 2 of T1 transgenic plants.The signals were detected on T2 plants from the other T1 transgenic plants.There were plants with SNR more than 2 from 4 plants of T1 transgenic plants.All these T2 plants with SNR more than 2 were positively transgenic plants by PCR.This study can jump over a series of in vitro molecular detection which is complex,time-consuming and costly,eliminates a large number of non-transgenic and poorly expressed plants or lines easily,in real-time,low-cost,intuitively.It provides a more efficient method for screening transgenic plants.