Banerjee Suchitra, Shang Tanya Q, Wilson Angela M, Moore Allison L, Strand Stuart E, Gordon Milton P, Lafferty Doty Sharon
Department of Biochemistry, Box 357350, University of Washington, Seattle, Washington 98195, USA.
Biotechnol Bioeng. 2002 Feb 15;77(4):462-6. doi: 10.1002/bit.10151.
P450 2E1 is an important mammalian liver enzyme known to metabolize a wide range of compounds including several common environmental pollutants. The medicinal plant, Atropa belladonna, was transformed with Agrobacterium rhizogenes containing a binary vector with rabbit P450 2E1 in either the sense or antisense orientation. The resulting "hairy roots" were isolated and grown in liquid medium. Production of P450 2E1 protein was verified in the roots containing the 2E1 gene in the sense orientation. Transgenic and control root cultures were dosed with the environmental pollutant, trichloroethylene (TCE), and were analyzed for the TCE metabolites, chloral and trichloroethanol. The root cultures expressing the mammalian P450 2E1 had increased levels of the metabolites compared to the levels in the control roots. This method represents a quick way to screen transformants for expression of foreign genes before regeneration of whole plants, and also as a possible source of foreign protein for purification.
细胞色素P450 2E1是一种重要的哺乳动物肝脏酶，已知它能代谢多种化合物，包括几种常见的环境污染物。药用植物颠茄用含有二元载体的发根农杆菌进行转化，该二元载体带有正向或反向的兔细胞色素P450 2E1。将得到的“毛状根”分离出来并在液体培养基中培养。在含有正向2E1基因的根中证实了细胞色素P450 2E1蛋白的产生。用环境污染物三氯乙烯（TCE）处理转基因根培养物和对照根培养物，并分析TCE代谢产物氯醛和三氯乙醇。与对照根中的水平相比，表达哺乳动物细胞色素P450 2E1的根培养物中代谢产物的水平有所增加。这种方法是在全植株再生之前筛选转化体中外源基因表达的一种快速方法，也是作为纯化外源蛋白的一个可能来源。
