CAJ | 학술논문

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-g-DNA 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.
재린산염완충용액중용1-(3-이갑안기병기)-3-을기탄이아알(EDC)여 N-간기호박선아알(NHS)활화최갑기섬유소납(CMC)측련상적최기；재실온하재장활화적 CMC 여5’단경안기수식적단련탈양핵당핵산(DNA)제취물(ODNs)반응,획득 CMC 상접지 ODNs 적공취물(CMC-g-ODNs),이 Lambda DNA 위모판,통과취합매련식반응(PCR),장접지적 ODNs 확증위장도위1300개감기대적쌍련 DNA,종이제득 CMC 측련상접지 DNA 적공취물 CMC-g-DNA.채용부리협홍외광보의측정 CMC 여 NHS 형성적중간체；용수평식경지당응효전영화수직판변성취병희선알응효전영대 CMC-g-DNA 접지공취물진행표정.결과표명,합성료 CMC-g-DNA 접지공취물,차재산성조건하 CMC 적활화효과경호；동시,접지재 CMC 상적 DNA 재경지당응효전영중천이속솔가쾌,이재취병희선알응효전영중천이속솔감만.
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-ethylcarbo-diimide(EDC) and N-hydroxy-succinimide(NHS) in phosphate buffer at both neutral and acid condition, re-spectively. 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 poly-merase 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 aga-rose gel electrophoresis, while it migrated slower than control polyacrylamide gel electrophoresis.