【摘要】 从安达盐碱地采收的蒙古柳种子在1/2MS固体培养基中培养,等到长出3～4片真叶的幼苗为实验材料,提取植株总RNA后分离mRNA,用SMART （Switching Mechanism At 5’end of RNA Template）方法合成蒙古柳cDNA,并连接到PYES₂载体,用电转化法将重组质粒转化到大肠杆菌感受态细胞JM109。经测定,原始文库滴度为6×10⁶cfu·m L^-1,重组率大于98%。插入片段的长度大部分在0. 5～2. 0 kb,平均长度为0. 8 kb,构建了蒙古柳c DNA-pYES₂大肠杆菌文库。更多还原

【Abstract】 In this experiment,the seeds of Mongolian willow harvested from Anda saline land were cultivated in 1/2 MS solid medium. Taking seedlings with 3 to 4 true leaves as experimental samples,the total RNA of the plant was extracted and the mRNA was isolated. The cDNA of Mongolian willow was synthesized by SMART method and ligated into PYES₂ vector. The recombinant plasmid was transformed into E. coli competent cell JM109 by electric transformation method.The original library titer was determined to be 6 × 10⁶ cfu·m L^-1,and the recombination rate was greater than 98%.The length of the insert is mostly 0. 5-2. 0 kb,with an average length of 0. 8 kb,and the cDNA-pYES₂ E. coli library of Mongolian willow was constructed.更多还原