CMC-g-DNA接枝共聚物的合成与表征  

Preparation and Characterization of the CMC-g-DNA Hybrid Copolymers

作　　者：李敏[1] 邢朝晖[1] 田艳芝[1] 姜勇[1] 

机构地区：[1]东南大学化学化工学院,南京211189

出　　处：《高等学校化学学报》2014年第7期1590-1595,共6页Chemical Journal of Chinese Universities

基　　金：国家自然科学基金(批准号:21174029);教育部留学回国人员科研启动基金资助~~

摘　　要：在磷酸盐缓冲溶液中用1-(3-二甲氨基丙基)-3-乙基碳二亚胺(EDC)与N-羟基琥珀酰亚胺(NHS)活化羧甲基纤维素钠(CMC)侧链上的羧基;在室温下再将活化的CMC与5'端经氨基修饰的单链脱氧核糖核酸(DNA)齐聚物(ODNs)反应,获得CMC上接枝ODNs的共聚物(CMC-g-ODNs),以Lambda DNA为模板,通过聚合酶链式反应(PCR),将接枝的ODNs扩增为长度为1300个碱基对的双链DNA,从而制得CMC侧链上接枝DNA的共聚物CMC-g-DNA.采用傅里叶红外光谱仪测定CMC与NHS形成的中间体;用水平式琼脂糖凝胶电泳和垂直板变性聚丙烯酰胺凝胶电泳对CMC-g-DNA接枝共聚物进行表征.结果表明,合成了CMC-gDNA接枝共聚物,且在酸性条件下CMC的活化效果更好;同时,接枝在CMC上的DNA在琼脂糖凝胶电泳中迁移速率加快,而在聚丙烯酰胺凝胶电泳中迁移速率减慢.Sodium carboxymethyl cellulose grafted DNA （CMC-g-DNA） hybrid copolymer, which can be a potential gene drug, was synthesized and characterized by both chemical and biological methods. First, the carboxyl groups on the side chain of CMC were activated by both 1-（3-dimethylaminopropyl）-3-ethylcarbodiimide（EDC） and N-hydroxy-succinimide（NHS） in phosphate buffer at both neutral and acid condition, respectively. After that, the CMC-NHS reacted with amino-functionalized oligodeoxynucleotides（ NH2-ODNs） to get CMC-g-ODNs grafted copolymer. Then CMC-g-ODNs were used as forward primers for the next-step polymerase chain reaction（PCR） to generate the final CMC-g-DNA hybrid copolymer. The infrared spectrum de- tection proved that the side carboxyl groups of the CMC were substituted very well by NHS and the efficiency was higher at acid condition than neutral. The obtained CMC-g-DNA copolymers were characterized by agarose gel electrophoresis and polyacrylamide gel electrophoresis, respectively. And the results showed that DNA was grafted on CMC successfully. An interesting finding is that CMC-g-DNA moved faster than control DNA in agarose gel electrophoresis, while it migrated slower than control polyacrylamide gel electrophoresis.

关 键 词：脱氧核糖核酸(DNA) 羧甲基纤维素钠 接枝共聚物 凝胶电泳 聚合酶链式反应 迁移速率 基因载体 

分 类 号：O636.9[理学—高分子化学]