DNAzymes inhibit bacterial TEM-1 β -lactamase gene expression in experimental study.docVIP

 PAGE \* MERGEFORMAT 2 DNAzymes inhibit bacterial TEM-1 β -lactamase gene expression in experimental study Author: Liu Gangling Baodong Xieyong En Lin Zhao Tingkun Lei Jun [Abstract] Objective To observe the 10-23 deoxyribozyme on bacterial enzyme TEM-1 gene expression in vivo. Methods PCR amplification of TEM-1 enzyme-wide coding genes, to connect into the pBK-CMV vector and expressed in E. coli JM109. Were designed and synthesized for the blaTEM-1 of the 10-23 DNAzyme, antisense oligonucleotides, using calcium chloride method to the respective expression of blaTEM-1 into Escherichia coli, the observation of bacteria into deoxyribozyme, the containing ampicillin (100μ g/ml) in the growth medium vitality and β -lactamase activity changes. Results 10-23 DNAzyme into the E. coli expression of blaTEM-1 after containing ampicillin (100μ g/ml) of the medium activity was significantly lower than the growth of Escherichia coli antisense oligonucleotide and blank the control group, into Escherichia coli deoxyribozyme β -lactamase activity is also significantly lower than that of antisense oligonucleotide and blank control group. Conclusion 10-23 deoxyribozyme can selectively inhibit bacterial blaTEM-1 enzyme gene expression. [Keywords:] resistance; β -lactamase; deoxyribozyme; gene expression ABSTRACT Objective To investigate the inhibitory effects of 10-23 deoxyribozyme (10-23 DRz) on blaTEM-1 gene expression in E.coli. Methods blaTEM-1 gene was amplified by polymerase chain reaction (PCR), the target amplification fragment was cloned into PBK-CMV vector. The recombinant vector was sequenced by Sanger’s dideoxy chain termination composition method. According to the gene sequence of blaTEM-1, 10-23 DRz and antisense oligonucleotides (As-ODN) were designed, synthesized, and introduced into E.coli producing TEM-1 respectively by the CaCl2 procedure, bacterial viability in liquid medium containing ampicillin (100μ g/ml) and β -lactamase activity were detected spectrop