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- W2355831454 abstract "Objective To construct DNA fusion fragments for metalloreductase gene disruption in Aspergillus fumigatus by SOE PCR,and determine the effects of nested PCR on SOE PCR.Methods The 5′-region and 3′-region of metalloreductase gene sequences were analyzed using sequence alignment and regions with high homology identified.Primers were designed in these regions to amplify the selected 5′-region and 3′-region of metalloreductase gene.At the same time,two DNA fragments were synthesized by PCR in hygromycin resistance marker cassette.By using corresponding DNA fragments as a template,fusion DNA fragments were amplified by nested PCR.The fusion DNA fragments were analyzed by gel electrophoresis and sequencing.Results The sizes of fusion DNA fragments were correct according to gel electrophoresis results.And it was verified that the fusion DNA fragments were linked correctly and were in high similarity with designed sequences by sequencing analysis.In contrast with general SOE PCR,the association of nested PCR reduced the amount of incorrect sized bands.Conclusion The DNA fusion fragments for metalloreductase gene disruption in Aspergillus fumigatus are obtained,which provide the foundation for disruption and investigation of metalloreductase gene function.Nested-SOE PCR can increase the specificity of PCR products." @default.
- W2355831454 created "2016-06-24" @default.
- W2355831454 creator A5074905453 @default.
- W2355831454 date "2013-01-01" @default.
- W2355831454 modified "2023-09-26" @default.
- W2355831454 title "APPLICATION OF NESTED-SOE PCR TO CONSTRUCTION OF DNA FUSION FRAGMENTS FOR METALLOREDUCTASE GENE DISRUPTION" @default.
- W2355831454 hasPublicationYear "2013" @default.
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