构建了一个CMV+GFP+EF1+目的基因的载体,当基因为mcherry时,cherry与GFP亮度一致约为85%左右,而当目的基因为OCT4/SOX2等多能性目的基因时,效率不到10%,求助大神们,这是为啥呀?
而当目的基因为OCT4/SOX2等多能性目的基因时,效率不到10%: how is this 10% rate determined? GFP or mCherry are fluorescent reporter genes, and are thought not to cause the testing cell gene transcription activity changed significantly, while OCT4 or SOX2 transgene expression could cause testing cell transcription activities changed markedly. To determine transgene expression efficiency DO NOT normalize fluorescent signals when OCT4/SOX2 transgene expressed with the fluorescent signals when only GFP or mCherry expressed. Try to run RNA sequencing to determine what the consequences caused by transgene OCT4/SOX2 expression.
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谢谢,意思是不要考虑GFP亮度,因为其转录活性可能被OCT4等影响了,直接检测OCT4的mRNA表达量? |