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Cloning and expression of Taxus acyltransferase cDNA
Jun Tu1, Ping Zhu, Ke-Di Cheng
1Institute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College, 1 Xian Nong Tan Street, Beijing 100050, P. R. China.
Abstract:
A new full-length acyltransferase cDNA was obtained from Taxus chinensis by homology-based cloning strategy. The cDNA has an open-reading frame of 1,275 nucleotides, which encodes 425 amino acids with a calculated molecular weight of 47,241 Da and an estimated pI value of 5.93. The deduced amino acid sequence resembles the sequences of other cloned acyltransferases (56-61% identity; 71-75% similarity) involved directly in taxol biosynthetic pathways. This cDNA was expressed in Escherichia coli using the expression vector pET32a(+). The expression band corresponds to the calculated mass plus the N-terminal fusion protein derived from the vector.
