用作植物疫苗的转基因胡萝卜的分子生物学筛选与鉴定

Screening and identification of transgenic carrot for plant vaccines

以包含有不同外源病毒基因的转基因胡萝卜为试验材料,应用分子生物学手段,从DNA水平上和蛋白质水平上对这些材料加以鉴定,经PCR引物筛选后得到转基因胡萝卜的阳性率为38%左右;为保证实验的可靠性和可行性,防止检测过程中遗漏和错误的发生,用特异的PCR引物对检测结果进行复选,得到的植株阳性率为7%.在蛋白水平上应用ELISA检测法检测获得的阳性植株的结果与PCR检测结果相同,且同时发现外源基因在胡萝卜根中的表达量明显小于在叶中的表达量.

Carrot was used as one of the bioreactor to produce vaccine. Several groups of primers were employed to detect the target DNA fragments. The results of PCR showed that the positive ratio reached to 38 % and 7 % respectively,by the universal primers and the special primers. ELISA method was also employed to detect the activity of expression of target gene. The results were matched with that of PCR. Amount of target gene expression in root showed less than that in the leaf.