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Chemcial Industry and Engineering 2015, Vol. 32 Issue (1) :59-63,78    DOI: 10.13353/j.issn.1004.9533.2015.01.012
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Cloning the Flanking Sequences of bapt by Thermal Asymmetric Interlaced PCR
Lin Chunyan, Guo Jing, Zhao Guangrong
Key Laboratory of Systems Bioengineering, School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, China

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Abstract Taxol is a natural anticancer drug produced from Taxus species, and widely used in the clinical treatment of breast cancer and small cell lung cancer. The expression of baccatin Ⅲ-aminophenylpropanoyl-13-O-transferase (bapt) gene is limited for taxol biosynthesis, and its flanking sequences are prerequisite for regulating taxol production. We cloned bapt gene and its flanking sequences with the length of 5.6 kb by homologous cloning and thermal asymmetric interlaced PCR. Bioinformatic analysis of the sequence showed that bapt gene contains two exons and one intron with the open reading frame of 1 338 bp. 5' Flanking sequence of 1 604 bp contains the promoter and 3' flanking sequence of 2 582 bp contains 242 bp long terminator. The regulatory elements discovered in this research would be helpful to improve the production of taxol through metabolically engineering taxus cells.
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Keywordstaxus;   taxol;   thermal asymmetric interlaced PCR;   baccatin Ⅲ-aminopheny;l propanoyl-13-O-transferase;   3-amino-3-phenyl-propanoyltransferase     
Received 2013-04-27;
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Lin Chunyan, Guo Jing, Zhao Guangrong.Cloning the Flanking Sequences of bapt by Thermal Asymmetric Interlaced PCR[J]  Chemcial Industry and Engineering, 2015,V32(1): 59-63,78
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