近日，福建省農(nóng)業(yè)科學(xué)院食品科學(xué)與技術(shù)研究所發(fā)表文獻(xiàn)《Whole-pathway stimulation of anthocyanin biosynthesis by UDP-glycose: flavonoid glycosyltransferase gene (UFGT) providing new insights into natural product development based on plant cell factory》，文獻(xiàn)中實(shí)驗(yàn)使用了LUYOR-3415RG便攜式雙波長熒光蛋白激發(fā)光源用于觀察GFP在愈傷中的表達(dá)，用于篩選轉(zhuǎn)基因陽性的細(xì)胞。

文獻(xiàn)摘要:

Detection of positive transformed cell lines

The resistance cell lines were observed using a fluorescent protein hand-held lamp (LUYOR-3415, Shanghai, China). The cell lines exhibiting green fluorescence characteristics were preliminary identified as positive transformed cell lines. These positive cell lines underwent further screening for three generations, with decreasing resistant concentrations, and were subsequently transferred to solid MS supplemented with 1.0 mg/L 2, 4-D without antibiotics for further subculture. Genomic DNA was extracted for PCR and sequencing to verify the transformation. And subcellular localization observation of UFGT was conducted using a FV1200 confocal microscope (Olympus, Tokyo, Japan).

3.2. Characteristics of transgenic cells and screening of high-yield anthocyanin cell line

Transgenic screening identified four positive grape cell lines (Fig. 2A), which exhibited a green fluorescent signal that diminished or disappeared with extended culture duration due to cellular senescence. Therefore, a portable fluorescent excitation light facilitates comprehensive full-stage observation and screening of transformed cells in vivo. These four positive grape cell lines were further validated using DNA-PCR (Fig. 2B). Subcellular localization analysis revealed that VdUFGT was localized in both the cytoplasm and nucleus (Fig. 2C), suggesting its potential role in the process. Both the WT and four VdUFGT transgenic cell lines exhibited varying degrees of red color (Fig. 2D). The transgenic cell lines generally displayed a deep red color on the surface, while the WT appeared light red, indicating a higher accumulation of anthocyanin in the transgenic cell lines. The proliferation coefficient results showed no significant differences among the different cell lines (Fig. 2E), suggesting that the overexpression of VdUFGT did not have a significant impact on the growth of grape cells. The anthocyanin content in the overexpressed cell lines was significantly higher than that in WT cell line, with the vgt3 cell line reaching 195.2 μg/g (FW), a 4.2-fold increase compared to the WT (Fig. 2F). However, the flavonoid content in vgt3 was significantly lower than that in WT cell line (1504.6 μg/g FW) (Fig. 2G). This discrepancy may be attributed to anthocyanins being terminal metabolites while flavonoids are intermediate ones. Consequently, the overexpression of VdUFGT promoted anthocyanin accumulation, with the vgt3 identified as a high-yield cell line for further utilization.

文獻(xiàn)地址：https://doi.org/10.1016/j.fbio.2024.105409 

我公司生產(chǎn)有手持式單波長熒光手電筒LUYOR-3430、LUYOR-3280,還有雙波長熒光蛋白激發(fā)光源LUYOR-3415、體視顯微鏡熒光適配器LUYOR-3421等一系列產(chǎn)品，具體產(chǎn)品介紹請瀏覽《GFP激發(fā)光源》，我公司提供樣機(jī)免費(fèi)試用，具體請按照網(wǎng)頁底部聯(lián)系方法電話或微信聯(lián)系。