Assessment of transfection by using cyanophycin and polyethylenimine cross-link with different cross-linking reagents

• Main Authors: Wen-hsuan Chen, 陳文玄
• Other Authors: Wen-chi Tseng
• Format: Others
• Language: zh-TW
• Published: 2013
• Online Access: http://ndltd.ncl.edu.tw/handle/32007716219036990631

碩士 === 國立臺灣科技大學 === 化學工程系 === 101 === Cyanopycin can be produced by recombinant Escherichia coli. It is a biodegradable polymer that consists of aspartic acid, arginine and lysine. The primary amine of lysine is easier to be chemically modified than the functional groups of arginine. In this study, we utilized the primary amine of lysine and low molecular weight polyethylenimine (PEI) to be cross-linked by different cross-linking reagents (glutaraldehyde, glucose, and genipin). The cross-linking reagents can react with primary amines. We expect that the cross-linked products can be used to transfect cells effectively. By DNA electrophoresis, we found that the cross-linked products by glutaraldehyde, glucose, and genipin have the ability to encapsulate DNA. We further transfected mammalian cell to examine the expression of green fluorescent protein. The results of transfection of Chinese Hamster Ovary (CHO) cells were observed under a fluorescent microscope. The cross-linked products by glucose resulted in a higher expression level of green fluorescent protein in comparisons with the other products by glutaraldehyde and genipin. However, the transfection efficiencies of those cross-linked products are not as good as that of high molecular weight PEI. The MTT assay was used to investigate the cellular viability after transfection. The results also showed that the cross-linked products by glucose are nontoxic.