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对农杆菌介导法获得的转 β_葡糖醛酸酶 (β_glucuronidase ,GUS)基因 (uidA)烟草 (NicotianatabacumL .)进行GUS表达分析 ,发现部分转基因植株无GUS活性。进一步Southern杂交结果发现 ,GUS基因失活植株的基因组中整合了多个uidA拷贝 ,而GUS活性高的转基因植株多为uidA单拷贝整合 ,表明uidA基因失活与基因多拷贝整合有关。Northern杂交结果显示 ,失活植株无特异uidARNA杂交带 ,而GUS活性高的植株可检测到明显的杂交信号 ,说明多拷贝引起的基因失活发生在RNA水平。
GUS expression analysis of β-glucuronidase (uidA) tobacco (Nicotiana tabacum L.) obtained by Agrobacterium tumefaciens-mediated method revealed that some transgenic plants had no GUS activity. Further Southern hybridization results showed that multiple copies of uidA were integrated into the genome of inactivated GUS plants, while those with high GUS activity were mostly single-copy uidA integration, indicating that inactivation of uidA gene was associated with multiple copies of genes. Northern hybridization results showed that the inactivated plant had no specific uidARNA hybridization band, while the plants with high GUS activity could detect obvious hybridization signals, indicating that multi-copy gene inactivation occurred at RNA level.