三种ARS片段对水稻部分酶解小细胞团转化频率的影响

ＡＲＳ２，ＡＲＳ７，ＲＳ８是从水稻珍汕９７Ａ不育系统ＤＮＡ中的克隆的一组在酵母细胞中能直始ＤＮＡ复制的片段。其中ＡＲＳ８是单考贝而ＡＲＳ２，ＡＲＳ７却是高度重复顺序。将带有ＡＲＳ２，ＡＲＳ７，ＡＲＳ８的质粒用低压脉冲电泳法导入水稻部分酶解小细胞团中，经浅层培养，铺板，获得愈伤组织。

The Effects of Three ARSs on the Transformation Frequence of Rice Partially Digested Small Group Cells

ARS2,ARS7 and ARS8 are a set of DNA fragments which are isolated from Zhenshan 97A rice male-sterile strain nuclear DNA and can initiate DNA replication in yeast. Among them, ARS8 is a single-copied DNA sequence in rice genome, while ARS2, ARS7 are highly repetitive DNA sequences. We introduce the ARS2,ARS7,ARS8 into rice partially digested small group cells mixture by low-voltage pulse field electrophoresis. After 30 days' incubation, transformed cells are plated onto the solid medium, and the callus was formed. After kanamysin screening and DNA extraction, Southern Dot boltting demostrates that 3. 2% of ARS2 transformed callus, 3. 6% of ARS7 transformed callus contain the foreign DNA. It is about 1 and 1. 3 times hight than the control YIP5-km respectively. Further works of Southern blotting reveals the pattern of multiple intergration site of ARS2 and ARS7.