新型荧光蛋白的从头合成及性质鉴定

Up to now the method to get fluorescent proteins (FPs) variants is focused on random mutation and site-directed mutagenesis. Herein we can not forecast and get the special expected mutated site at our will for random mutation. Though site-directed solved this problem, its inherent limitation that it is difficult to get amount mutation at the same time and it is a money-spending and time-consuming method hindered its application. In this article we used a high throughput and relatively cheap technology- microfludic PicoArray method for the simultaneous de novo synthesis fluorescent proteins（FPs）DNA sequences and reassemble sequences successively to get massive variants of fluorescent proteins. The result showed that five FPs are unique and have properties different to the others FPs existed and within these five new FPs, three FPs with different hues are belong to red fluorescent proteins which emission wavelength are longer than 550 nm and the other two locate at green fluorescent protein region. The native gel combined with Mass spectrum showed that all the five FPs are monomer with molecular weight are between 25 kD and 35 kD and they have relative stronger fluorescence intensity comparing with former blue fluorescent proteins, so they will be potential tools in molecular and medical studying fields.