Preparation and Characterization of the CMC-g-DNAHybrid Copolymers†

doi:10.7503/cjcu20140045

Abstract

Abstract:

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, 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 detection 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.

Key words: DNA, Sodium carboxymethyl cellulose(CMC), Hybrid copolymer, Gel electrophoresis, Polymerase chain reaction(PCR), Migrated speed, Gene combination

CLC Number:

O636.9

TrendMD:

LI Min, XING Zhaohui, TIAN Yanzhi, JIANG Yong. Preparation and Characterization of the CMC-g-DNAHybrid Copolymers^†[J]. Chem. J. Chinese Universities, 2014, 35(7): 1590.

Figures/Tables 5

Scheme 1 S ynthetic routes of CMC-g-DNA

Fig.1 Polymerase Chain Reaction(PCR) as a tool for the preparation of CMC-g-DNA

Fig.2 FTIR spectra of CMC(a), activated CMC at pH=7.2(b) and activated CMC at pH=6.5(c)

Fig.3 Picture of 3% horizontal agarose gel electrophoresis of 1000 bp DNA Marker P(lane 1), unreacted ODNs(lane 2), CMC-g-ODNs(lane 3, CMC activated at pH=7.2) and CMC-g-ODNs(lane 4, CMC activated at pH=6.5)

Fig.4 Picture of 1% horizontal agarose gel electrophoresis(A) and 5% vertical polyacrylamide gel electrophoresis(B) (A) Lane 1: 1000 bp DNA Ladder; lane 2: 1300 bp DNA; lane 3: CMC-g-DNA; lane 4: CMC-g-ODNs. (B) Lane 1: 1300 bp DNA; lane 2: CMC-g-DNA.

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