子房滴注法将GUS基因表达框转入大豆的研究

Transfer of a GUS gene Cassette into Soybean via Ovary-drip Transformation

为了验证大豆子房滴注转化方法的可重复性与遗传稳定性,将由报告基因GUS、表达调控序列(35S启动子,NOS终止子)和T-DNA边界序列3部分组成的基因表达框转入大豆。对T0代转化材料进行GUS组织化学染色分析和PCR检测。结果表明:在检测的340个幼胚中有12个呈GUS阳性,且在180个转化植株中有6个呈PCR阳性同时其叶片也呈GUS阳性,转化率分别为3.53%和3.33%。Southern杂交表明外源GUS基因表达框以低拷贝的形式整合到大豆基因组中。对转基因后代叶片GUS染色、PCR分析及Northern blot检测结果表明外源基因已遗传给了后代,其中S2株系符合孟德尔分离规律。

To validate the reproducibility and genetic stability of the ovary-drip transformation,the GUS gene cassette,which was composed of only the GUS gene, expression regulatory, sequence （ 35S CaMV promoter, NOS terminator） , and T- DNA border sequence at both sides, was introduced into soybean. The results（ Histochemistry GUS analysis and PCR screening of T0generation） showed that 12 out of 340 detected immature embryo were GUS positive and six out of 180 transformed plants were PCR positive and GUS positive,the rate of the T0 generation transformation reached 3.53% and 3.33% , respectivly. Southern blot analysis confirmed that GUS gene was integrated into the soybean genome with low copy numbers. The GUS staining,PCR analysis and Northern blot indicated that GUS gene was inherited in the transgenic progenies. Moreover,Mendelian segregation was observed in the S2 line.