摘要
利用PCR技术扩增了酿酒酵母(Saccharomyces cerevisiae)Ⅲ号染色体上以ARS(Autonomously replicating sequence,ARS)305为核心的不同长度侧翼序列,并将其构建到酵母整合型载体pRS405中获得了重组质粒PA500、PA1000、PA2000,然后通过电转化法转入酿酒酵母细胞中,测定了不同ARS305片段对酵母转化效率及质粒在酵母中稳定性。结果表明,并非侧翼序列越长ARS活性越高,重组质粒PA1000具有比PA2000更高转化频率和转化稳定性,推测ARS两侧序列存在一些顺式和反式的调控机制影响ARS自主复制功能。
Autonomously replicating sequence （ARS）305 gene and its flanking sequence of Saccharomyces cerevisiae were cloned by PCR,and then connected into the shuttle vector pRS405.Recombinant plasmids PA500,PA1000,PA2000,were successfully constructed and transformed by electroporation to S.cerevisiae,which determine transformation efficiency for different fragments of ARS305 and plasmid stability.The results demonstrated that ARS activity did not have a positive correlation with the length of the flanking sequences,and PA1000 was higher than PA2000 in the frequency of transformation and conversion stability.It suggested that ARS activity was affected by flanking sequence that has some cis-and trans-regulatory mechanism.
出处
《湖北农业科学》
北大核心
2013年第19期4804-4806,4822,共4页
Hubei Agricultural Sciences
基金
国家自然科学基金项目(61072129)
内蒙古自治区自然科学基金项目(2011MS0504)
