乳腺癌T7噬菌体展示cDNA文库的构建  

Construct breast carcinoma T7 phage display cDNA library

作　　者：张涛[1] 施宝民[1] 王洪[2] 陈鹊汀[2] 季堃[1] 余松林[1] 

机构地区：[1]同济大学附属同济医院普外科,上海200065 [2]河北大学附属医院肿瘤外科,保定031000

出　　处：《外科研究与新技术》2015年第1期9-11,共3页Surgical Research and New Technique

摘　　要：目的构建乳腺癌组织T7噬菌体展示c DNA文库,为下一步筛选差异蛋白打下基础。方法利用乳腺癌新鲜标本,提取总RNA,分离m RNA并进行纯化,然后合成c DNA,连接体外包装获得T7噬菌体展示c DNA文库。结果总RNA经检测,A260/A280=1.87,纯化的m RNA产量为4.0μg,A260/A280=1.91,合成的c DNA大小在200~6 000 bp之间,原始文库的容量为2×10^7pfu,文库重组率为90%,插入片段长度在300~2 000 bp之间。结论噬菌体展示技术是进行蛋白质功能研究的高效方法,构建高质量的乳腺癌噬菌体展示c DNA文库,可用于肿瘤标志物的筛选、肿瘤疫苗的研制、多肽药物的开发、靶向治疗的研究等众多领域。Objective To construct the breast carcinoma T7 phage display c DNA library,so the foundation for the screening of differentially expressed proteins was laid. Methods Firstly,fresh specimens of breast carcinoma was used to extract total RNA,separating and purifying of m RNA. Then synthesis c DNA was done. At last,the packaging was connected in vitro and T7 phage display c DNA library was obtained. Results The total RNA was tested,the result is A260/A280=1.87. The purified m RNA is 4.0 μg,A260/A280=1.91. The size of c DNA is 200- 6 000 bp. The primary library capacity is 2× 10^7 pfu. Recombination rate of the library is 90%. The length of inserted fragments is 300- 2 000 bp. Conclusions Phage display is an efficient method of protein function research. We constructed a high quality breast cancer phage display c DNA Library. The library can be used for screening tumor markers,tumor vaccine,polypeptide drug development,targeted research,and so on.

关 键 词：乳腺癌 噬菌体展示 c DNA文库 

分 类 号：R737.9[医药卫生—肿瘤]