转多基因杨树中抗生素标记基因NPTⅡ表达量分析研究

Expression Analysis of Neomycin Phosphotransferase Ⅱ(NPTⅡ) Gene in Multigenes Transgenic Poplar

对转基因库安托杨及银腺杂种杨（含1-5个外源基因）中选择标记基因新霉素磷酸转移酶基因（NPTII）进行PCR扩增,证实该基因稳定存在于转基因杨树的基因组中。应用酶联免疫吸附测定法（ELISA）对2种转基因杨树中NPTII蛋白的表达量进行定量检测,结果显示：NPTII蛋白的表达量并没有随着目的基因数量的增加而升高,说明与转单个基因杨树相比,转多个基因杨树可能由标记基因引起的生物安全问题并不会加剧。

PCR amplification was applied on selectable marker gene neomycin phosphotransferase II （NPTII） in two transgenic poplar species, P. x euramericana＇ Guariento＇ and Populus alba x P. glandulosa which contain one to five foreign genes. The results showed that this gene was stable integrated in the genome of poplar trees analyzed. To quantify the expression of NPTII gene, enzyme linked immunosorbent assay was performed on both transgenic poplar species. The results showed highly significant linear correlation between quantity of NPTII proteins and their OD values, while the quantity of NPTII proteins expressed in different transgenic lines did not increase with the increased number of transgenes in both poplar species. Our results indicated that the potential biosecurity risk caused by muhiplegenes transgenic poplar was not more serious than that of single gene transgenic poplar.