SMARTer技术构建辣椒黄绿苗突变体叶片全长cDNA文库  

Construction of full-length cDNA library of yellow bud mutant leaves in Capsicum annuum L.using SMARTer technique

作　　者：马志虎[1] 孙国胜[2] 张昌伟[2] 杨玉霞[2] 潘跃平[1] 

机构地区：[1]江苏丘陵地区镇江农业科学研究所,江苏句容212400 [2]南京农业大学园艺学院,江苏南京210095

出　　处：《江苏农业学报》2013年第2期389-393,共5页Jiangsu Journal of Agricultural Sciences

基　　金：江苏省自然科学基金项目(BK2012698)

摘　　要：本研究以辣椒黄绿苗嫩叶为材料,提取总RNA,采用LD-PCR技术合成First-strand cDNA和ds cDNA。将分级纯化后的ds cDNA连接到载体pSMART2IFD上,用电穿孔法将重组子转化到大肠杆菌感受态细胞DH5α中,构建辣椒全长cDNA文库。文库质量检测结果显示:原始文库滴度为1.76×106PFU/ml,重组率为94%,插入片段长度为500～2 000 bp,平均长度为1 170 bp,表明构建的辣椒叶片cDNA文库较为理想,可用于目的基因筛选。Total RNA was extracted from yellow bud mutant leaves of Capsicum annuum L. , and first-strand cDNA and ds cDNA were synthesized by LD-PCR technology. The purified ds cDNA was connected to vector pSMART2IFD, and the recombinant vectors were transformed into competent Escherichia coli cells DH5α by electroporation to construct fulllength cDNA library of Capsicum annuum L. The library quality test results showed the titer of original library was 1.76× 10^6PFU/ml, the recombination rate was 94%, and the inserted fragment length was 500-2 000 bp, indicating that the library was ideal for target genes selection.

关 键 词：辣椒 黄绿苗突变体 SMARTer CDNA文库 

分 类 号：S641.3[农业科学—蔬菜学]