人促红细胞生成素基因在番茄中的表达

Expression of Erythropoietin Gene in Tomato

通过子叶与农杆菌（ＡｇｒｏｂａｃｔｅｒｉｕｍｔｕｍｅｆａｃｉｅｎｓＡＧＬ１）共培养，将表达载体ｐＨＬＰＥ中的人促红细胞生成素（ｈＥＰＯ，一种人来源的典型糖蛋白）基因（ｅｐｏ）导入番茄，然后用卡那霉素进行筛选，获得了抗性植株。经点杂交和Ｓｏｕｔｈｅｒｎ印迹分析，证明部分抗性植株中整合了ｅｐｏ基因。通过对转基因番茄植株叶片的粗提蛋白进行ＥＰＯＥＬＩＳＡ检测，结果表明，ｈＥＰＯ在番茄中的表达量为约４００ｐｇ／ｇ叶片。经用转基因植株叶片粗提蛋白饲喂依赖于ＥＰＯ的ＴＦ１细胞，表明番茄ＥＰＯ具有体外（ｉｎｖｉｔｒｏ）生物活性，这暗示用植物生产的ＥＰＯ可作为体外药物加以应用。

? The plant expression vector pHLPE contained hEPO cDNA under control of the cauliflower mosaic virusderived 35S RNA promoter and NOS terminator was constructed and was introduced into Agrobacterium tumefaciens strain AGL1 by direct transformation. By using the modified leafdisc cocultivated method, hEPO cDNA was transferred into tomato cells. Integration of the cDNA into the genome of tomato plants was confirmed by Dot blot and Southern blot analysis. hEPO protein was shown to be expressed in tomato plants by colorimetric enzyme immunoassay. The productivity of EPO in the transformed tomato plants T302 and T505 was 444 and 342pg/g of leaf respectively. Tomatoproduced EPO exhibited in vitro biological activities in the assay with TF1 cell, which suggests that EPO produced in tomato could be used as in vitro pharmaceuticals.