摘要
目的构建用于酵母双杂交的HL-60细胞ds cDNA文库。方法采用CLONTECH公司提供的SMART技术，提取对数生长期HL-60细胞的总RNA，逆转录生成第一链cDNA，LDPCR合成dscDNA，与pGADT7-Rec共转化酵母菌AH109。结果成功构建了用于酵母双杂交的HL-60细胞ds cDNA文库，AD转化效率为8×10^4cfu／3μg p GADT7-Rec。结论HL-60细胞ds cDNA文库可以充当酵母双杂交系统中的cDNA文库，用于进一步筛选与M—CSF相互作用的非受体靶分子。
Objective To construct ds cDNA of HL-60 cells in yeast two-hybrid sys- tem. Methods With the SMART technology supplied by CLONTECH Company, the total RNA of HL-60 ceils was extracted and reversed to form the first cDNA, and ds cDNA was then synthesized through LD PCR. The ds cDNA was co-transformed with pGADT7-Rec into AH109 strain. Results The ds cDNA of HL-60 cells was successfully constructed in yeast two-hybrid system. AD co-transfor- mation efficacy reached up to 8×10^4 cfu/3μg pGADT7-Rec. Conclusion The ds cDNA of HL-60 cells can act as cDNA library in yeast two-hybrid system for screening the target molecules interac- ting with M-CSF.
出处
《医学分子生物学杂志》
CAS
CSCD
2012年第2期84-87,共4页
Journal of Medical Molecular Biology
基金
国家自然科学基金（No.30270684）,湖南省自然科学基金重点项目（No.09JJ3060）,湖南省教育厅科技计划重点项目（No.10A087）,湖南高校科技创新平台基金（No.09K072）
