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- W2363350809 abstract "The overlap extension mediated by restriction endonuclease to obtain the full length gene was established. Two partial fragments were isolated separately. A proper restriction site was cleavaged by the corresponding restriction endonuclease, and the two fragments were overlapped, used as template for each other and extended into a full length gene under catalysis of DNA polymerase. The exon 9 of cyclooxygenase 1 (COX 1) gene was inserted into COX 1 cDNA fragment without exon 9, and the full length cDNA of COX 1 gene was obtained. The method includes three steps. Firstly, the partial COX 1 cDNA fragment containing exon 9 and COX 1 cDNA without exon 9 were isolated separately by RT PCR, and cloned into pMD18 T vector. Secondly, the partial COX 1 cDNA containing exon 9 was amplified by PCR, and was mixed with combinative plasmid without exon 9 cleaved by Stu Ⅰ. The two fragments used as template for each other, and overlap extended into a double strand DNA molecule under catalysis of pfu DNA polymerase. Finally, the full length cDNA of COX 1 gene containing exon 9 was obtained by PCR using the terminal primer of COX 1 cDNA. The overlap extension mediated by restriction endonuclease was especially applied for cloning genes regulated on mRNA splicing level,and also expected to be a new method for gene recombination and modification." @default.
- W2363350809 created "2016-06-24" @default.
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- W2363350809 date "2004-01-01" @default.
- W2363350809 modified "2023-09-22" @default.
- W2363350809 title "Application of Overlap Extension Mediated by Restriction Endonuclease in Cloning of Human Cyclooxygenase-1 (COX-1) Gene" @default.
- W2363350809 hasPublicationYear "2004" @default.
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