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KMID : 0545120100200111481
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Journal of Microbiology and Biotechnology
2010 Volume.20 No. 11 p.1481 ~ p.1483
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Construction of a T-vector using an esterase reporter for direct cloning of PCR products
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Lim Ho-Dong
Cheong Dae-Eun
Shin Hyun-Jae
Kim Geun-Joong
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Abstract
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We constructed an efficient T vector pTQEST216T that employed an engineered esterase as an indicator for direct cloning of PCR products. After ligation of the XcmI-digested vector with PCR products, this cloning system could easily discriminate positive clones due to insertional inactivation of the esterase reporter. Additionally, PCR products were cloned into this vector efficiently without the gel purification steps, due to the well-designed multi-cloning site that was in-frame fused at the circularly permutated gap of the reporter.
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KEYWORD
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Esterase, Reporter, TA cloning, T-vector, PCR product
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