35SP-rolB基因在棉花转基因系基因组中转录稳定性研究

Transcriptional Stability of 35SP-rolB Gene in Transgenic Cotton(Gossypium hirsutum L.) Genome

以8个T7代的棉花35 SP-rolB转基因系为材料,通过提取转基因株系及对照植株叶片总DNA和总RNA,对各个转基因株系分别进行PCR检测和Northern杂交,研究转基因的遗传稳定性和转录稳定性。结果表明,35 SP-rolB目的基因和NPTⅡ选择标记基因在经过对转基因系的连续多代定向选择后,在各个转基因系基因组中都能够稳定遗传和高效转录,说明针对强生根、早发、抗早衰等特殊性状进行定向选择是保持rolB转基因系遗传稳定性和转基因表达稳定性的有效途径。

Eight 35SP-rolB transgenic lines of T7 generation of upland cotton were applied to analyze the stabilities of transgenice inheritance and transcription.Total DNA and RNA were prepared from the leaf extracts of the rolB transgenic lines and the control,and the result was verified by PCR amplification and Northern hybridization.The results showed that the 35SP-rolB gene and the NPTⅡ selection marker in the genomes of transgenic lines,which were selected for seven successive generations based on the specific traits of strong rooting ability,decrepitude resistance and rapid recovery after seedling transplantation,behaved stable inheritance and transcription in all eight transgenic lines.The results also demonstrated that the orientated selection at toward target traits is an efficient way in keeping the genetic stability of the transgenic lines and the expressional stability of the 35SP-rolB transgene.